AGRI-FOOD XXV - ULB Sibiusaiapm.ulbsibiu.ro/cercetare/conferinte/AFSPT2015/ProcFood.pdf · Proceedings of the Conference with International Participation “AGRI-FOOD SCIENCES, PROCESSES

“Lucian Blaga” University of Sibiu Faculty of Agricultural Sciences, Food Industry and

Environmental Protection

ORGANISE THE CONFERENCE WITH INTERNATIONAL

PARTICIPATION

“AGRI-FOOD SCIENCES, PROCESSES AND TECHNOLOGIES”

AGRI-FOOD XXV

Celebrating the XXVth Anniversary of the Beginning of Food Industry and Agronomy

Higher Education in Sibiu

CONFERENCE SECTION

Food

Sibiu,

May 24-26, 2015

Published in Romania by ”Lucian Blaga” University of Sibiu ISSN 1843-0694

Proceedings of the Conference with International Participation “AGRI-FOOD SCIENCES, PROCESSES AND TECHNOLOGIES”

AGRI-FOOD XXV, May 24-26, 2015, Sibiu, Romania

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Scientific Committee Prof. Eng. Constantin-Horia BARBU, Ph.D. - “Lucian Blaga” University of Sibiu, Romania Prof. Eng. Petre ALEXE, Ph.D. - “Dunărea de Jos” University of Galaţi, Romania Prof. Eng. Petruţa CORNEA, Ph.D.- University of Agricultural Sciences and Veterinary Medicine, Bucharest, Romania Prof. Eng. Maya IGNATOVA, Ph.D. - Academy of Sciences, Sofia, Bulgary Prof. Eng. Dumitru MNERIE, Ph.D. - Polytechnic University of Timişoara, Romania Prof. Simona OANCEA, Ph.D. - “Lucian Blaga” University of Sibiu, Romania Prof. Eng. Laszlo SIKOLYA, Ph.D. - College of Nyregyhaza, Hungary Prof. Eng. Anca ŞIPOŞ, Ph.D. - “Lucian Blaga” University of Sibiu, Romania Prof. Eng. Dumitru ŢUCU, Ph.D. - Polytechnic University of Timişoara, Romania Assoc. Prof. Adriana BIRCĂ, Ph.D. - Technical University of Moldova, Moldova Assoc. Prof. Maria CSUBAK, Ph.D. - University of Debrecen, Hungary Lecturer Mihaela ANTOFIE, Ph.D. - “Lucian Blaga” University of Sibiu, Romania Lecturer Eng. Olga DRĂGHICI, Ph.D. - “Lucian Blaga” University of Sibiu, Romania

Organizing Committee Prof. Eng. Ovidiu TIŢA, Ph.D.- Dean Prof. Eng. Camelia SAVA, Ph.D.– Director of SAIPA Department Lecturer Eng. Ioan Dan MIRONESCU, Ph.D. – Vice-dean Prof. Eng. Gligor Ciortea, Ph.D. Prof. Neli Darie, Ph.D. Prof. Eng. Ec. Romulus Iagăru, Ph.D. Prof. Eng. Mihaela Tiţa, Ph.D. Assoc. Prof. Eng. Robert Blaj, Ph.D. Assoc. Prof. Iuliana Bratu, Ph.D. Assoc. Prof. Monica Creţu, Ph.D. Assoc. Prof. Eng. Mariana Dumitru, Ph.D. Assoc. Prof. Eng. Monica Mironescu, Ph.D. Assoc. Prof. Maria Ioana Moise, Ph.D. Assoc. Prof. Eng. Victor Nederiță, Ph.D. Assoc. Prof. Eng. Maria Tănase, Ph.D. Assoc. Prof. Eng. Adrian Turtureanu, Ph.D. Assoc. Prof. Eng. Ioan Vonica, Ph.D. Lecturer Iuliana Antonie, Ph.D.

Lecturer Eng. Iulian Bratu, Ph.D. Lecturer Eng. Cristina Anca Danciu, Ph.D. Lecturer Cecilia Georgescu, Ph.D. Lecturer Eng. Lia Iagăru, Ph.D. Lecturer Eng. Maria Lidia Iancu, Ph.D. Lecturer Eng. Ramona Iancu, Ph.D. Lecturer Eng. Otto Kettney, Ph.D. Lecturer Eng. Ecaterina Lengyel, Ph.D. Lecturer Eng. Cristina Moise, Ph.D. Lecturer Eng. George Moise, Ph.D. Lecturer Eng. Claudia Ognean, Ph.D. Lecturer Eng. Mihai Ognean, Ph.D. Lecturer Petronela Pavel, Ph.D. Lecturer Eng. Mihai Pop, Ph.D. Lecturer Eng. Ioan Pășcănuț, Ph.D. Lecturer Eng. Mariana Liliana Păcală, Ph.D. Lecturer Eng. Mircea Savatie, Ph.D. Lecturer Daniela Simtion, Ph.D. Lecturer Simona Spânu, Ph.D.



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Distingueshed guests, dear colleagues and students,

I have the great pleasure to welcome you on the Transylvanian realms, at the Faculty of Agricultural Sciences, Food Industry and Environmental Protection of Sibiu. Every time we see each other in these situations, there is a feeling of emotion and pleasure, generated by the need of knowing and of establishing of camaraderie relationships, beneficial to human development and professional achievements. Today we celebrate 25 years of continuity for the higher education in food industry at Sibiu, continuing a more than 70 years one in Romania. It is a beautiful evolution and a confirmation of the teaching and research quality within our Faculty, which may be seen and taken for granted, but in fact encompasses a lot of work, dedication and even sacrifice.

It has been, during human evolution, a permanent quest for improving the standard of living, mainly by food quality and diversification, and this will hold as long life will exist on Earth. Soil and plant agronomy will enable the obtaining of raw materials in accordance with the actual quality requirements and their industrial processing will define and ensure the right nutrition, also respecting their traceability. Romania has always had exquisite conditions for obtaining raw materials with the highest quality which, carefully processed, have provided an excedent of the domestic, then being a “hard currency” for Romanian exports. Now there are good conditions for obtaining organic food in Romania, but these conditions should be more exploited in our own benefit, to contribute more and to enlarge the national GDP. These are facts that, corroborated with the scientific potential of our researchers and manufacturers’ practical expertise can make the difference in competiveness within a society in continuous move of the areas of influence and food markets.

The Faculty of Agricultural Sciences, Food Industry and Environmental Protection intents to stay connected to the European requirements and standards, by educating specialists able to adjust themselves to the advanced system of technological and scientific values. Situated in a city with old cultural traditions and historical memories, it pursues the work of carving and shaping the human and spiritual values, giving a new dimension to scientific knowledge, in a climate of normality. By its team of academic staff, students and doctoral candidates we have managed to keep steady our Faculty among the first ones within the Lucian Blaga University of Sibiu in what concerns scientific research and obtaining European funding, of paramount importance for structuring a successful education activity, comparable with other European similar institutions.



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Nowadays we have as priority providing a favourable climate for creativity and novelty of higher education in agronomy and food industry, by encouraging research and teaching, for the integration into the world scientific community. Within this context we have created and developed study programs that are interested for the license, master and doctoral students, with the appreciation of industrial stakeholders.

The year 2015 has an exquisite importance for the academic community of Sibiu, being a celebration year for our University that has since 20 years as spiritual father a multivalent personality of Romanian culture, the poet and philosopher Lucian BLAGA. It is a happy destiny that our faculty shares with the university and local community we belong to.

I would like to thank in this way to all that during the past time has contributed to the creation of the present of us all – colleagues, students and partners – for the edification of the future. I wish you all success, health and happiness.

With esteem and friendship,

Prof. dr. Ovidiu TIȚA



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SUMMARY

Authors

Paper’s title Pag.

1 Mariana Popescu, Florin Oancea, Carmen Lupu, Elena Radu, Mălina Deșliu-Avram

BY-PRODUCTS FROM WINE LEES HYDRODISTILLATION WITH POTENTIAL USE FOR FEED SUPPLEMENTS

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2 Ioana-Codruta Racz

POOR DIET – A DRIVER OF GROWTH DISORDERS IN CHILDREN

19

3 Camelia Rovinaru, Diana Pasarin, Malina Desliu

ADAPTATION OF A NON-CONVENTIONAL YEAST STRAIN WHICH METABOLISE LACTOSE BY ESTABLISHMENT OF CULTURE MEDIA COMPOSITION BASED ON WHEY

26

4 Butoi Simona, Butoi Laurențiu, Tiţa Mihaela,

Prăvariu Ileana

RESEARCH ON THE CHARACTERISTICS OF OBTAINING YOGURT FROM COW’S MILK AND SOY MILK

35

5 Diana Ionela Stegăruș, Ecaterina Lengyel, Ovidiu Tița, Ciucure Corina Teodora

IMPROVING GENETIC RESOURCES OF MICROORGANISM’S VINEYARDS IN ORDER TO OBTAIN A HIGH QUALITY WINES, VINEYARDS FROM LOCAL AREAS

40

6 Adrian Turtureanu COMPARATIVE STUDY ON NICKEL REMOVAL FROM AQUEOUS SOLUTIONS BY FLOTATION WITH ANIONIC AND CATIONIC COLLECTOR

47

7 Neli Darie

NATURAL ANTIOXIDANT FEATURE ESTIMATED IN NON-ALCOHOLIC DRINKS BY CHEMILUMINOMETRY

55

8 Carmen Monica Crețu

RESEARCHE REGARDING ANODIC OXIDATION OF ALUMINIUM FROM ACID MIXTURE ELECTROLYTE

60

9 Daniela Maria Şandru

BIOACTIVE PLANTS - VALUABLE SOURCE OF TANNINS

66

10 Iancu Maria Lidia, Iancu Bianca Maria

USE OF LOCAL NATURAL RESOURCES AT IMPROVE THE QUALITY PREMIXES PRODUCT FOR GFD

70

11 Otto Ketney

DECONTAMINATION KINETICS OF AFLATOXIN M AT DIFFERENT DOSES OF UV RADIATION

76

12 Ecaterina Lengyel PHYSICAL AND GEOGRAPHICAL PARTICULARITIES OF VINEYARDS IN THE APOLD DEPRESSION

84



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13 Mihaela Amăriuţei,

Mihaela-Adriana Tiţa, Simona Oancea

EVALUATION OF ASCORBIC ACID AND TOTAL ANTHOCYANINS IN FRESH CHEESE ENRICHED WITH BILBERRY, CRANBERRY AND DRIED BERRIES

90

14 Ramona Maria Iancu

AUTHENTICITY OF DAIRY GOAT MILK EXISTING IN ROMANIAN MARKET

97

15 Mihai Ognean, Olga Drăghici, Maria Lidia Iancu

COMPARISON BETWEEN TWO INSTRUMENTS FOR EVALUATION OF DOUGH RHEOLOGY DURING KNEADING

103

16 Elena-Roxana Tufeanu, ŞtefanOancea, Monica Mironescu, Ion Mironescu, OvidiuTiţa

RHEOLOGY OF WAXY MAIZE STARCHES AS TOOL FOR INDUSTRIAL APPLICATIONS

111

17 Cristina Anca Danciu

ADJUSTMENT OF CORRUGATIONS NUMBER/CM, FOR THE BREAKING ROLLERS, RELATED TO THE GRINDING RESISTANCE OF THE WHEAT GRAIN

117

18 Mihaela Balteș

IDENTIFICATION AND CHARACTERIZATION OF USEFUL SUBPRODUCTS OF GRAPEWINE PRODUCTS, CASE STUDY: WINE YEAST

125

19 Natalia Țislinscaia, Mircea Bernic, Andrei Lupaşco, Alexei Buleandra, Iana Ţislinscaia, Răducan Marcel, Mihai Melenciuc

MOIST MATERIALS DRYING PROCESS ENERGY CONSUMPTION CALCULATION

131

20 Mircea Bernic, Natalia Ţislinscaia, Lupaşco Andrei, Vasile Cartofeanu, Marcel Răducan, Leonid Zavialov, Mihai Melenciuc M.

DRYING KINETICS OF ALMOND SEEDS USING PULSED UHF ENERGY INTAKE

137

21 Ciprian Căpăţână, Mihai Văduva

DETERMINATION OF RELATIVE SPEED OF PARTICLES IN LIQUID FLOWING IN TUBES AT HIGH TEMPERATURES

145

22 Ciprian Căpăţână REDUCEING HUMIDITY LOSSES OF FRUIT PIECES BY COATING WITH BEESWAX

148

23 Adina Frum EXTRACTION OF ANTHOCYANINS FROM BY-PRODUCTS FROM THE WINEMAKING PROCESS

152



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BY-PRODUCTS FROM WINE LEES HYDRODISTILLATION WITH

POTENTIAL USE FOR FEED SUPPLEMENTS

Mariana Popescu*, Florin Oancea*, Carmen Lupu**, Elena Radu*, Mălina Deșliu-Avram*

* National R&D Institute for Chemistry and Petrochemistry ICECHIM Bucharest, Romania

**R&D Institute for Plant Protection ICDPP Bucharest, Romania

Abstract

Wine lees, the by-product after grape must fermentation, contain mainly dead yeasts and other materials

deposited on the bottom of the fermentation vessels. This solid waste from decanting maturated and clarified

wines should be completely valorized to enhance wineries competitivity, ecoefficiency of the technology and

wine quality. Agri-food domains are the most suitable to use the products obtained by wine lees conversion in

several applications as fertilizers, antioxidants, aromatic additives or emulsifiers in organic agricultural

practice, animal feeding, beverages and food industries. Hydrodistillation is a simple and clean process

consisting in heating a water suspension of lees, separating the fractions and identifying the valuable

constituents. This research work is a comparison between several experimental models based on the

hydrodistillation principle, using two different laboratory equipments and various technological parameters.

Yeast cell proteins, fermentation glycerol, ethyl esters of fatty acids, tartaric acid and sugars were identified as

by-products of wine lees hydrodistillation and proposed as potential natural raw materials for new value

added (bio)products such as feed supplements in modern ecological formulations.

Keywords: hydrodistillation, wine lees, spent yeast, tartaric acid

INTRODUCTION

Valorization of agroindustrial wastes and by-products into edible added value products

represents an international challenge for developed society to protect environment and

improve economical performance of nations. Winemaking is one of the dominant agro-

industrial activities in Europe (Salgado et al., 2014) being one of the polluting sectors since

estimated 14.5 million tons of grape by-products had been yearly produced (Chouchouli et al.

2013). Wine production in European Union is over 153.771.000 hl, Italy, France and Spain

being the main countries producing wine in Europe. Romania has also a long tradition of

grape wine production from small wineries to large enterprises. Continental climate and soil

quality are suitable for viticulture development on many regional agricultural areas.



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International recognition of several varieties of wines from Romanian wineries stimulated the

intensification of efforts to improve the quality and quantity of wine production every year.

As a consequence, large quantities of by-products and wastes accumulated and producers

must find technical solutions for conversion of spent biomass into acceptable products for

their integrated technological flow or for commercial purposes in accordance with the

international legislation concerning environmental protection.

The nature and physic-chemical properties of the residual material are closely

dependent on the specific winery procedures and their characteristics determine further

applications and specific valorization circuit to be integrated as raw materials. Innovative

solutions must be proposed and tested to develop a real sustainable industry (Devesa et al.,

2004).

Wine lees are one of the most undervalued by-products of winemaking industry.

“Wine lees is a residue that forms at the bottom of recipients containing wines, after

fermentation, during storage or after authorized treatments, as well as residue obtained

following the filtration or centrifugation of this product” (EEC regulation No. 337/79). Wine

lees representing almost 14% of organic waste after alcoholic fermentation is a permitted

source for alcohol production according to the European Council Regulation (EC) 479/2008.

Lees must be collected in containers and delivered to suitable authority having the respective

permission for transferring to a distillery for the commercial production of ethanol (Naziri et

al., 2014) and should not bee discharge on the field being harmful (Moldes et al., 2008) and

incompatible with agricultural requirements because of heavy metals contain and low pH 3,8-

6,8 (Bustamante et al., 2008). Wine lees compositions depends on grape variety, major

constituents being microorganisms (mainly yeast cells of Saccharomyces cerevisiae), tartaric

acid (Naziri 2012), inorganic matter and phenolic compounds (Pérez-Serradilla et al., 2011).

Wine lees are sources of functional lipid squalene, an all-trans linear triterpenoid hydrocarbon

with great importance for food industry, pharmaceuticals and cosmetics (Naziri et al., 2014).

During wine aging, autolysis of the yeast cell membrane release hydrolytic enzymes which

break intracellular biopolymers into smaller molecules. Lees seems to have a poor nutritional

value as an animal nutrient (Rivas et al., 2006), however, yeast cell membrane rich in lipids

releasing (poly)unsaturated fatty acids (oleic, linoleic and linolenic) represent a potential

supplement for human and animal feeding (Gomez et al., 2004)

Winery by-products are separated from the lees through distillation in stainless-steel

stills. Major products are bioethanol and aromas responsible for organoleptic properties of



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wines. After ethanol distillation, lees can be used for the recovery of tartaric acid to be reused

in wine production (Rivas et al., 2006). DL-Tartaric Acid is a mineral supplement, most

commonly used in nutritional supplement and food additive applications. Essential oils, also

known as wine lees essence, are widely used in the pharmaceutical, fragrance, cosmetic,

flavoring and wellbeing industries and are obtained through hydrodistillation of the lees.

Hydrodistillation is a variant of steam distillation in which material is soaked for some time in

water after which the mixture is heated and volatile materials are carried away in the steam,

condensed and separated. This is the standard method for extracting essential oils for quality

control of vegetable products. During hydrodistillation, most of the essential oils form

azeotropic mixtures with water, being separated and decanted from the condensation liquid

after 30 minutes or longer time of heating. Temperature of exposed material must be generally

around 100ºC to avoid decomposition of products, elevated pressure should be applied for

plants whose essential oils are difficult to extract and reduced pressure being better when

lower temperatures are recommended.

This research work presents three experimental models for hydrodistillation of wine

lees performed at laboratory level with different experimental parameters in order to identify

by-products with potential use as ingredients for feed supplements.

MATERIALS AND METHODS

EQUIPMENTS

♦ Red wine lees from year 2014 production in Romanian Dobrogea region Murfatlar

♦ Enzymatic extract Glucanex (Novozymes)

♦ Distilled water

♦ NaOH analytical reagent (SC Chimreactiv SRL)

♦ Clevenger glass apparatus for light oils (SC Adrian System SRL)

♦ Laboratory glass installation for simple distillation

♦ Ph-meter

♦ Büchner vacuum tube with filter papers

♦ Laboratory glassware (funnels, plates, sticks)

♦ Electric grinder

♦ CaCl2 desiccators

♦ LabSwift-aw portable (Novasina)



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EXPERIMENTAL MODELS

Three experimental models were applied using hydro-distillation principle: material to be

extracted was immersed in water and boiled, the vapors of the volatile components were

carried by the steam to a condenser and a liquid was collected. The first variant of the

experimental models was performed in a Clevenger apparatus for light oils using crude wine

lees and the others in a laboratory installation for simple distillation using enzimatically

hydrolyzed and crude lees respectively, applying mild vacuum and temperature in order to

avoid decomposition of valuable by-products. Samples of wine lees were distilled to remove

ethanol and extracted with water, treated with calcium acetate to precipitate calcium tartrate

and acidified with HCl to tartaric acid. Fractions of distillate were separated and their

chemical composition was assayed with analytical GC/MS and NISTMS library for the

identification of the compounds. Spent yeast was air dried and grinded, samples being

collected and placed in CaCl2 desiccators. Water activity (aw) of yeast dried powder was

periodically analyzed using a LabSwift-aw portable device to evaluate the storage stability of

this substrate depending on the resistance to biological contamination and being correlated

with the relative humidity (rh): rh (%) = aw x 100

♦ Variant 1

A portion of 50 grams of wine lees and 200 ml distilled water were placed into Clevenger

apparatus for light oil extraction. Aqueous lees suspension was boiled with reflux a period of

3.5 hours at high temperature (200°C) on thermoelectric bath. Recycled distillate was

collected, spent yeast was filtered with light vacuum and the liquid was used at the second

step by mixing with a new portion of crude lees in weight ratio of about 2:1. The distillation

was repeated as for the first step, collecting the recycled distilate, filtering the spent yeast and

using the liquid to the third step by mixing it with another portion of crude lees in weight ratio

of 1:1. Two unrecycled portions of distillation heads were collected (about 30 ml), spent yeast

was filtered and dried. The final filtrate was concentrated by simple distillation to precipitate

crystals of tartaric acid and salts. This experimental model will be called „hydrodistillation by

waterfall process”.

♦ Variant 2

A portion of 100,58 gr wine lees (pH=3,61) was suspended in 150 ml distilled water, and

treated with concentrated water solution of NaOH to pH=6,2 and a concentrated aqueous

enzimatic extract Glucanex 0,5 g under magnetic stirring (380 rpm), 1 hour at 30°C and 3

hours at 60°C. Enzymatically hydrolyzed lees were izolated from the suspension by



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centrifugation and samples from recovered wet biomass were dried (105°C) and assayed for

water activity determination. A portion of 238 ml hydrolyzed yeast was placed into a round

1.5 liter bottle with two necks equipped with magnetic stirrer, a thermometer and a glass

descendent cooler. The mixture was heated using an oil thermostatic bath, applying a reduced

pressure of 250-300 mmHg and the distillate were collected into glass vessels. Bottoms were

filtered under light vacuum, yeasys were washed with distilled water and dried, and the

filtrate was concentrated near dryness by light vacuum distillation.

♦ Variant 3

A portion of 150 grams of crude red wine lees and 150 ml distilled water were placed into a

round 1.5 liter bottle with tree necks equipped with a thermometer, a glass funnel, and a glass

descendent cooler. The mixture was heated using an oil thermostatic bath, with magnetic

stirring and reduced pressure of about 300 mmHg and the fractions were similarly processed

as in the variant 2.

RESULTS AND DISCUSSIONS

Hydrodistillation in three steps waterfall process resulted in three portions of distillates, three

portions of spent yeast and the final filtrate which was concentrated by simple distillation to

obtain the fractions presented in Table 1.

Table 1. Material balance in hydrodistillation waterfall process

*samples assayed for chemical composition

Final filtrate produced by concentration near dryness 82 ml distillate and 72 grams bottoms

with pH=4 which decanted 26 grams of brown syrup and a wet precipitate producing after

drying 4 grams of crystals. Another portion of 8 grams of crystals was obtained after washing

with water the spent yeast and concentrating the filtrate. Finally, 37 grams of dry spent yeast

Step no.

Input Output

Crude lees Liquid Distillate Filtrate

Wet spent yeast

Dry spent yeast

Loss

g mL Type mL pH mL pH g g g

1 50 200 water 17 4.0 200 3.5 14 4.55 19

2 100 190 Filtrate from 1st step 16 4.5 192 4.0 74 14.95 8/10*

3 180 180 Filtrate from 2nd step 30 4.0 212 4.0 98 26.50 20/12*

Total 330 200 63 212 186 46 47/22*



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were obtained after 24 hours at the ambiental temperature having water activity aw= 0,222

(relative humidity rh=22,2%) and after 24 hours in a CaCl2 dessicator aw =0,123 (rh=12,3%)

was detected at 22,9ºC with portable LabSwift-aw.

Chemical composition of the fractions is presented by the most representative gas-

chromatografic diagrams (GC chromatogramms).

Figure 1 – Chemical composition of the recycled distillate from the first step of

the waterfall hydrodistillation of red wine lees in Clevenger apparatus for light oils

In the first step, the distillate was an azeotropic mixture (Figure 1) of water with the

major component bioethanol (97.99%), traces of ethyl acetate (0,16%), fermentation amyl

alcohol (1,09%), ethyl lactate (0,21%) and ethyl succinate (0,21%).

Figure 2 – Chemical composition of recycled distillate from the second step of the

waterfall hydrodistillation of red wine lees in Clevenger apparatus for light oils



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In the second step, the same components from the new portion of lees were obtained

and several ethyl esters of fatty acids appeared (Figure 2): ethyl octanoate (0,15%), ethyl

decanoate (0,47%), ethyl dodecanoate (0,34%), ethyltetradecanoate (0,09%).

In the third step, the unrecycled head of distillation (the first 30 ml) had the

aproximately same composition as the fist step (Figure 1) with small differences of the

compound concentration: major component bioethanol (97.52%) and traces of ethyl acetate

(0.67%), fermentation amyl alcohol (1.38%), ethyl lactate (0.17%) and ethyl succinate

(0.26%). If the middle distillate was not separed from crude lees hydrodistillation, ethyl esters

of hexadecanoic (17,07%), octadecanoic (5,06%), oleic (11,14%), linoleic (17,4%), linolenic

(6,19%), eicosanoic (2,95%) acids and fermentation glycerol (31,03%) were recovered in the

dried bottom of the waterfall hydrodistillation process (Figure 3).

Figure 3. Chemical constituents (others than bioethanol) of the third dried

bottom from waterfall hydrodistillation of red wine lees in Clevenger apparatus

These valuable unsaturated fatty acid esters were not present in distillation heads, nor

in the crude lees composition (Figure 4).

The second and the third experimental models were performed in order to confirm the

yeast origin of fatty acids generating the ethyl esters by-products during the hydrodistillation

of wine lees.



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Figure 4- Chemical constituents others than bioethanol

from crude lees supernatant

Comparing the chemical compositions of supernatantes from centrifugated crude

(Figure 4) and enzimatically hydrolyzed lees, excepting the bioethanol, the release of fatty

acids from biological cells was demonstrated by the presence of several saturated and

unsaturated fatty acid ethyl esters (Figure 5).

Figure 5 - Chemical constituents others than bioethanol from enzimatically

hydrolyzed lees supernatant



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Hydrodistillation with mild vacuum and temperature conditions in laboratory

conventional installation for simple distillation in the second and third experimental models

using crude and enzimatically hydrolyzed wine lees resulted in the separation of four main

fractions: distillation head, middle distillate, bottoms and spent yeasts presented in Table 2.

Table 2 – Hydrodistillation fractions obtained with mild vacuum and temperature

Var. no.

Distillation head

Middle distillate

Bottoms Spent yeast Wet Dry Wet Dry

mL pH mL pH g g g g rh0 (%)

rh24 (%)

rh48 (%)

2 126 4.22 96 4.20 56 10 32 10 21.8 15.6 15.4 3 55 3.45 146 3.37 45 10 87 23 24.9 16.6 15.8

Mild conditions of hydrodistillation of crude or hydrolyzed lees gave similar results

for the composition of the distillation heads with those of recycled or nonrecycled distillation

heads from Clevenger apparatus for light oils since they did not favoured the extraction of

ethyl esters of organic acids with more than C4 (Figure 6).

Figure 6 - Middle distillate composition from the third experimental model

The valuable unsaturated fatty acid esters (Figure 3) which were not present in

distillation heads (Figure 1) nor in the crude lees composition (Figure 4) also were not



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evidenced in the middle distillate when mild condition of pressure and temperature were

applied (Figure 6).

Final processing of the solid bottoms produced a wet bottom which dried as a

glycerinous syrup and spent yeasts obtained as dry powders having the qualities requested for

natural substrates as evidenced by their relative humidity valors (Tabel 2).

Figure 7. Dry powder of spent yeast by-product from the three experimental models of red wine

lees hydrodistillation

Different parameters of the three experimental models generated different types and

properties of by-products such as purple to brown powder of dried spent yeasts (Figure 7) and

caramel to molasses flavored and colored crystals to jelly bottom by-products (Figure 8).

(a) (b)

Figure 8. By-products from the red wine lees hydrodistillation: (a) wet glycerinous bottom and (b)

dried crystals from concentrated syrups



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Red wine lees also contained 3,96% tartrates which correspond to 3,13% tartaric acid,

antocyanic pigments and 0,28 % polysaccarides based on the dried substance from lees which

hydrolyzed in smaller molecules of sugars also enriching the flavor and appearance of the

syrupy bottoms.

CONCLUSIONS

Wasteless agri-food industry represents a challenge for scientific researchers,

producers and international authorities in the future years to 2025. Valorization of by-products

and wastes has been considered the optimal solution by environmental and economical

reasons. Wastes should be used as raw materials in new products for various applications, the

out-put of one sector becoming in-put for other sector or recycled as defined by the modern

concept of integrated management for sustainable development of the economy. Winery

wastes are one of the most unexploited natural matters and unprocessed disposals are

dangerous for agroecosystems. Minimal processing should convert wine lees into valuable

ingredients for feed supplements or other ecological agri-food products.

Hydrodistillation is a simple and inexpensive clean technology for separation of

different fractions from natural raw materials without using organic solvents or other

chemicals and being suitable for experimental applications from laboratory to industrial level.

The main product of hydrodistillation was bioethanol which was collected in the heads

of distillation with traces of ethyl esters of acids C2-C4 and amyl alcohol. Experimental

model variants 2 and 3 demonstrated that mild conditions of pressure and temperature do not

favorized C8-C12 esters extraction by hydrodistillation. The composition of distillates and

bottoms depended on the wine lees preprocessing and the hydrodistillation process parameters

(temperature, time, pressure, water to lees ratios). One or another variant of hydrodistillation

and optimal operational parameters should be chosen as function of the desired by-products.

The ethyl esters of fatty acids C8-C12 identified in the middle distillate from crude

lees in the second step of waterfall process are known as flavors and aromatics for alcoholic

beverages. Fermentation glycerol, glucolipids from yeast cells and tartaric acid from bottom

fractions of hydrodistillation should act as natural additives, antioxidants and preservatives.

Spent yeast by-product should be used as an adsorbent matrix for active ingredients in feed

supplements having beneficial properties for health of animals and humans due to constitutive

proteins and unsaturated fatty acids released from their cellular walls.



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Future works of experimental research should complete and optimize the

hydrodistillation by waterfall process in order to raise the yields and quality of valuable by-

products: fatty acid ethyl esters, glycerinous syrup, tartaric acid and spent yeasts. These

preliminary results demonstrated the opportunity of valorization the by-products from

hydrodistillation of wine lees as ingredients for feed supplements.

AKNOWLEDGEMENTS

This research work has been performed within PNII Programme Partnerships in Prioritary

Areas, Project PNII-PT-PCCA-2013-4-0425, Contract 156/2014 financially supported by

UEFISCDI, Romanian Ministry of Education and Scientific Research

REFERENCES 1. Bustamante M.A., Moral R., Paredes C., Pérez-Espinosa A., Moreno-Caselles J., Pérez-Murcia M.D.,

Agrochemical characterization of the solid by-products and residues from the winery and distillery industry,

Waste Management, 2008, 28, 372–380.

2. Chouchouli V., Kalogeropoulos N., Konteles S.J., Karvela E., Makris D.P., Karathanos V.T., Fortification of

yoghurts with grape (Vitis vinifera) seed extracts, Food Science and Technology, 2013, 53, 522–529.

3. Devesa-Rey R., Vecino X., Varela-Alende J.L., Barral M.T., Cruz J.M., Moldes A.B., Valorization of winery

waste vs. the costs of not recycling, Waste Management, 2011, 31, 2327–2335.

4. Gómez M.E., Igartiburu J.M., Pando E., Rodríguez-Luis F., Mourente, G., Lipid composition of lees from

Sherry wine, Journal of Agricultural and Food Chemistry, 2004, 52, 4791-4794

5. Moldes A.B., Vásquez M., Domínguez J.B., Díaz-Fierros B., Barral M.T., Negative effect of discharging

vinification lees on soils, Bioresource Technology, 2008, 99, 5991-5996

6. Naziri E., Mantzouridou F, Tsimidou M.Z., Recovery of squalene from wine lees using ultrasound assisted

extraction, Journal of Agricultural and Food Industry, 2012, 60, 9195-9210

7. Naziri E., Nenadis N., Mantzouridou F., Tsimidou M.Z., Valorization of the major agrifood industrial by-

products and waste from Central Macedonia (Greece) for the recovery of compounds for food applications, Food

Research International, 2014, 65, 350-358

8. Pérez-Serradilla J.A., Luque de Castro, M.D., Microwave-assisted extraction of phenolic compounds from

wine lees and spray-drying of the extract, Food Chemistry, 2011, 124, 1652–1659.

9. Rivas B.,Torrado A., Moldes A.B., Domínguez J.M., Tartaric acid recovery from distilled lees and use of the

residual solid as an economic nutrient for Lactobacillus, Journal of Agricultural and Food Industry, 2006, 54,

7904-7911

10. Salgado J.M., Abrunhosa L., Venâncio A., Domínguez J.M., Belo I., Integrated use of residues from olive

mill and winery for lipase production by solid state fermentation with Aspergillus sp. , Applied Biochemistry and

Biotechnology, 2014, 172, 1832-1845



19

POOR DIET – A DRIVER OF GROWTH DISORDERS IN CHILDREN

Ioana-Codruta Racz

PhD Fellow, SOP HRD/159/1.5/S/133675 Project, “Lucian Blaga” University from Sibiu, Partner

Abstract

Proper nutrition is the most important determinant of health status of children. Healthy eating is the main

method to ensure proper height and weight growth. Among the micronutrients required for a satisfactory

nutrition, iodine plays an important role, outlines the main substrate in the formation of thyroid hormones,

involved in growth and development of the human body, starting with intrauterine life. Iodine deficiency is an

important public health problem, although various improvements have been made in this regard by universal

iodization of salt for human consumption. Assessment of nutritional status of children is done by determining

a series of anthropometric parameters, the most common being the weight and height, which are compared

with those of a reference population. In assessing short stature is commonly used standard deviation or

percentile, defining itself as height below -2 standard deviations from the average for the corresponding age

and gender.

Keywords: nutrition, children, iodine deficiency.

INTRODUCTION

Proper nutrition is one of the most important determinants of health status of children.

Normal somatic growth and mental development, and the social, economic and familiar level

are premises for sustainable development of the society. A healthy diet is the main method of

ensuring proper somatic growth, accompanied by adequate psycho-intellectual development, a

good functioning of the human body organs and systems, protection against harmful external

factors in the environment. The two major categories of nutrients from food, macronutrients

represented by proteins, fat and carbohydrates, and micronutrients including vitamins and

minerals, are essential to a healthy diet that provides adequate growth and development of the

body.

Nowadays, along with macronutrients, a particular attention has acquired micronutrients,

especially minerals. Among nutritional factors, an important role in child health, even in the

intrauterine life, are iron and iodine, which justifies the importance given their status in

pregnant women and in children.1 Iodine is an essential component in the body and plays an

important role in the growth and development of all organs. Iodine is essential for synthesis of



20

thyroid hormones, thyroxin (T4) and triiodothyronine (T3), hormones that are vital for normal

growth, and particularly for the developing of the brain and central nervous system.2

In 1980, WHO estimates that up to 60% of the world population was deficient in

iodine, mostly in developed countries. It’s been realized that the consequences of iodine

deficiency could be avoided by a low-cost intervention, universal iodization of salt for human

consumption.3 Since then, globally, the number of households using iodized salt has increased

from under 20% to over 70%, dramatically reducing the iodine deficiency.4 Despite this

enormous progress, in 2007, WHO estimated that nearly two billion people still have

insufficient iodine intake, including 1/3 children of school age.5

Unlike other nutrients, iodine does not occur naturally in some foods, rather, it is

present in the soil and it is ingested through food grown in that soil. Iodine deficiency results

from uneven distribution of iodine in the soil. The highest content of iodine is founded in the

sea fish or seafood. Although lower than in seafood, iodine content in meat is greater than in

vegetables, iodine content of food provided by animals may be enriched by the addition of

iodized salt in animal nutrition.1 Iodine deficiency in the soil can’t be corrected, so the food

grown on these soils do not provide enough iodine for the basic needs of the population. In

Romania there are several areas deficient in iodine, especially mountain regions, among

which are certain areas in Sibiu County.

According to the World Health Organization, the optimum iodine requirement of a

child of school age is 100-199 micrograms/day. In pregnant women, iodine requirement

increases by 50%, which mean 150-249 micrograms/day iodine intake.6

As we mentioned above, iodine is a micronutrient who takes part of the thyroid

hormones, being captured by the thyroid gland, where, by several processes form thyroid

hormones. These hormones have an essential role in cell differentiation during intrauterine

development and help to maintain heat homeostasis and metabolic function at adult age.7

Assessment of nutritional status is done by determining a series of anthropometric

parameters, the most common being the weight, height, body mass index and for

micronutrients, laboratory determinations from blood or biological products. In terms of

normal growth, children's height and weight is compared to that of a reference population

(healthy children of the same age and gender).1 In assessing height currently is used standard

deviation or percentile.

Standard deviation is a measure commonly used for dispersion from the mean of a set

of individual values. In case of a Gaussian distribution of the height of children with the same



21

age, race and gender, children with a size range from the mean +/- 1DS will be many, and

they can be considered to have a normal size for their age. If we enlarge the limit to +/- 2DS,

children with height between -1DS and -2DS will be small, but with a big chance of being

normal. Children who exceed the limit of +/- 2DS are rare, with a risk of manifesting a

pathological substrate for deviation from the average height, greater as the deviation is more

higher.8 In clinical practice, if we refer to small stature, any child with a height size below

-2DS than average corresponding to children with the same age, race and gender, is suspected

to be suffering from a disorder of growth, the degree of suspicion increasing inversely

proportional to the size of the child.8


The purpose of this paper is to assess weight status and growth in school children aged 6-14

years, students of 0 to VIII class, in an area of Sibiu County known as region with moderate

iodine deficiency, Gura Riului. We proposed to evaluate the prevalence of children under -

2DS (standard deviation) of the reference standards for height, and the prevalence of children

with changes in weight status, with obesity or malnutrition.

The following indicators were used to assess the growth and development: weight related to

height, reflecting the weight status of these children; height related to age, reflecting skeletal

development. For these determinations were used specific measurement instruments, like a

properly calibrated scale and a taliometer, the same for all measurements. Given that in our

country we do not have recent data on normal somatic values, the last dating from 1980, for

this work we used tables with standard deviations for age and gender of countries with similar

characteristics with our country on anthropometric normal values, respectively

Switzerland.9,10 All measurements were made after we obtained the permission of Sibiu

County School Inspectorate, and the permission of the parents and teachers of the school

visited.

RESULTS

398 children were evaluated, of which 194 (49%) were male and 204 (51%) female.

Depending on the height of children, they were divided into 3 groups: children who fall

within the normal range for age and gender of their height, so they have average height, or

height between +2DS and -2DS; the second group consists of those children who already

present small stature, height below -2 standard deviations, and a third group including



22

children with severe retardation of stature, having -3DS or less. Thus, 363 (91%) of the

children are within the normal range of height, 20 (5%) have small stature with height under

-2DS (mention that in this group were classified children with height of -2DS also) and finally

15 (4%) children have severely retarded stature (-3DS or less).

Fig.1. Distribution by height

If we refer to children with small stature between -2DS and -3DS, we can say that 40% of

them are female, 60% are male and of those with severe retardation of stature, below or equal

to -3DS, 7% are girls and 93% boys.

Table 1. Distribution by gender and height Normal height (-2DS, -3DS) ( <= -3DS)

Female 195 (54%) 8 (40%) 1 (7%)

Male 168 (46%) 12 (60%) 14 (93%)

Total 363 (100%) 20 (100%) 15 (100%)

Of the nine girls with short stature, one is aged 6 years, 3 girls are aged 7 years, 4 girls are

aged 8 years and the last one is 10 years old. Of the 26 boys with small stature 3 are aged 7

years, 11 are aged 8 years, 3 boys are 9 years old, 4 boys are 10 years old, two were aged 12

and the last 3 boys are 11, 13 and 14 years old.



23

Fig. 2. Short stature – distribution by age and gender

In terms of weight status, children were divided into four categories, namely those with

normal weight 350(88%), children with obesity degree I - 35(9%), grade II obese children -

5(1%) and children with poor weight status, malnutrition - 8(2%). Between children with

obesity degree I, a number of 19 are girls, 16 are boys; two girls and three boys presented

grade II obesity; 7 girls and a boy presents malnutrition.

Fig.3. Distribution by weight

Referring to the weight status of children with short stature, out of 35 children with growth

retardation, one child has obesity degree I and one child has malnutrition, the remaining 33

children have normal weight for their age and gender.

DISCUSSIONS

Repeated studies have shown that Gura Riului is a moderate iodine deficiency region in the

area of Sibiu County, the last study in 2009 demonstrated the persistence of iodine deficiency

even after the application of the strategy of universal salt iodization. After 5 years from the



24

last published data in this area, in 2009, iodine deficiency did not vary significantly,

confirming the need for specific supported prophylaxis.11,12 Worldwide, although they have

shown vast improvement in iodine status in the past decade, there are still areas where iodine

deficiency is present. "Progress has slowed in the past decade, is limited primarily by

technical problems to reach small producers of salt, weak quality control of iodized salt and

difficulties in enforcement of iodized salt" wrote Elizabeth N. Pearce as, MD associate

professor of medicine at the University of Boston, Massachusetts, USA.

Studies on iodine deficiency and somatic growth of children have been conducted in Europe

and beyond, showing different results. In Greece, school-age children in regions with endemic

goiter due to low iodine intake, showed height and weight lower than other children from a

non-endemic area of the same age and gender.14 Other similar studies in Iran and India

showed at children in iodine deficient regions of these countries, a growth retardation.15,16

Worldwide, progress targeting the iodine deficiency was enormous, since the main strategy of

universal iodization of salt for human consumption was adopted in 1993, in Romania since

2004. Today, iodine deficiency decreased, but there are still regions where iodine deficiency

is present, affecting mainly children and pregnant women, thus requiring further support to

continue the fight against this health problem.

CONCLUSIONS

The ratio male/female children examined was approximately equal, 49% were male and 51%

female.

Of all children evaluated, 9% of them had short stature, as 35 of 398 children.

The age group that we found most children with short stature was 8 years.

9% of all children have obesity degree I, and 2% have poor weight status (malnutrition).

Of the 35 children with growth failure found, only 2 children have a weight status changed,

the rest having normal weight.

ACKNOWLEDGEMENTS:

This paper is supported by the Sectoral Operational Programme Human Resources

Development (SOP HRD), financed from the European Social Fund and by Romanian

Government under the contract number POSDRU/159/1.5/S/133675



25

REFERENCES 1. UNICEF, Principii în alimentația copilului și a gravidei, Ediția a-2-a, Ed. MarLink, București, 2007.

2. Skeaff SA., Iodine Deficiency in Pregnancy: The Effect on Neurodevelopment in the Child, Nutrients, 2011,

3, 265-273.

3. Zimmermann MB., Jooste PL., Pandav CS., Iodine-Deficiency Disorders, Lancet, 2008, 372, 1251-1262.

4. Andersson M., de Benoist B., Rogers L., Epidemiology of iodine deficiency: salt iodisation and iodine status,

Best Pract. Res. Clin. Endocrinol. Metab., 2010, 24, 1-11.

5. de Benoist B., McLean E., Andersson M., Rogers L., Iodine deficiency in 2007: global progress since 2003,

Food Nutr. Bull., 2008, 29, 195-202.

6. WHO, UNICEF, International Council for Control of Iodine Deficiency Disorders, Assesement of iodine

deficiency disorders and monitoring their elimination: a guide for programme managers, Geneva, WHO, 3rd Ed.,

2007.

7. Melmed S., Polonsky KS., Larsen PR., Kronenberg HM., Williams Textbook of Endocrinology, 12th Edition.

Ed. Elsevier Sauders, Philadelphia, 2011.

8. Brănișteanu DD., Tulburările de creștere. Ghid de diagnostic și tratament, Ed. Polirom, Iași, 2011.

9. Romer T. et al., Efficacy and safety of a new ready-to-use recombinat human growth hormone solution,

Journal Endocrinology Invest, 2007, 30, 578-589.

10. Romer T. et al., Seven years of safety and efficacy of Recombinant Human Growth Hormone Omnitrope in

the treatment of Growth Hormone Deficient Children: results of a phase III study, Hormone Research, 2009, 72,

359-369.

11. Rusu N., Totoianu IG., Evoluția statusului iodat la școlarii din Gura Rîului (Sibiu) prin determinarea ioduriei,

Acta Medica Transilvanica, 2009, 2, 81-84.

12. Stanciu M., Totoianu IG., Evaluarea deficitului iodat prin determinarea ioduriei la școlarii din Gura Rîului,

Sibiu, Revista Sibiul Medical, 2003, 19, 1.

13. Neville AN., Iodine Deficiency Still a Concern, Despite Vast Improvements, Thyroid, 2013, 23, 523-528.

14. Koutras DA. et al., Endemic goiter in Greece: nutritional status, growth and skeletal development of goitrus

and nongoitrus population, Am. J. Clin. Nutr., 1973, 26, 1360-1368.

15. Azizi F., et al., Physical, neuromotor and intelectual impairement in non-cretinous school children with

iodine deficiency, Int. J. Vit. Nutr. Res., 1995, 65, 199-205.

15. Lal RB., Srivastava VK., Chandra R., A study of the spectrum of iodine deficiency disorders in rural area of

Uttar Pradesh, Indian J. Public Health, 1996, 40, 10-12.



26

ADAPTATION OF A NON-CONVENTIONAL YEAST STRAIN WHICH

METABOLISE LACTOSE BY ESTABLISHMENT OF CULTURE

MEDIA COMPOSITION BASED ON WHEY

Camelia Rovinaru, Diana Pasarin, Malina Desliu

Institutul National de Cercetare-Dezvoltare pentru Chimie si Petrochimie – ICECHIM

Abstract

In the context of this work we evaluated, in preliminary tests, aerobic growth of Kluyveromyces marxianus

ZIM 1867 as a producer of yeast biomass on different sweet whey media, containing mineral sources,

organic nitrogen source and yeast extract, for an active multiplication of yeast cells. The effect of medium

composition on the production of yeast biomass was investigated in shake flask culture. Among different

growth media compositions, the medium composed of enzymatic hydrolyzed sweet whey yielded the

highest percentage of dry matter of 5,06%, after 24 hours cultivation, at 280C and 150 rpm. The results

obtained showed that it is of considerable value if a safe microorganism likes K. marxianus can utilize

whey permeate lactose to generate profitable quantities of biomass, useful by-product.

Keywords: Kluyveromyces marxianus, whey, biomass, growth

Introduction

Media components have a very strong impact on economics of industrial

fermentation processes and can account for up to 30% of the total production cost (Rivas

B, 2004)

The composition of the medium used for cultivation of micro-organisms is directly

reflected in their physiological phenotype and their fermentation performance, which in

turn affects the results of strain analyses and strain performance in industrial applications.

Furthermore, economic constraints make the large-scale production of low-cost products

reliant on cheap sources of carbon and nitrogen, such as molasses from the sugar industry,

corn steep liquor from the starch industry, spent sulfite liquor from the forest products

industry and cheese whey from the dairy industry (Dahod SK, 1999)

Large scale production of cheap commodities such as fuels, chemicals and

materials requires very cheap raw material (Miller T, 1986, Dahod SK, 1999). Such

processes use by-products from the agricultural, forestry and chemical industry as carbon



27

and nitrogen sources. Carbon sources include sugar beet and sugar cane molasses, residues

from sugar production, spent sulfite liquor (SSL) from the paper pulping industry, and

cheese whey from the dairy industry.

Cheese whey is a by-product of dairy industries, that is formed during the

coagulation of milk casein in cheese making or in casein manufacture; represents about

85–95% of the milk volume and retains 55% of milk nutrients, such as: lactose (4.5–5%

w/v), soluble proteins (0.6–0.8% w/v), lipids (0.4–0.5% w/v), mineral salts (8–10% of

dried extract), lactic (0.05% w/v) and citric acids, non-protein nitrogen compounds (urea

and uric acid) and B group vitamins (Siso M, 1996).

This paper presents an alternative for whey valorization by culturing fungal strains

belonging to the genus Kluyveromyces, which include unicellular microorganisms, eukaryotic,

non-motile, and is widely used on an industrial scale because of its ability to aerobic

metabolize whey lactose with biomass accumulation. Many strains of the genus

Kluyveromyces are sources of β - galactosidase and are recognized as GRAS (General

Recognized As Safe) for industrial use (Dziekak, 1987).

The aim of this study is to investigate the effect of different whey media compositions

on Kluyveromyces yeast biomass growth and selecting the best medium option with good

biomass growth relative to dry substance, efficient both technological and economical.

Materials and methods

The fresh whey was obtained from dairy producing farms in Alba and preserved by

freezing to prevent lactose hydrolysis by Lactobacillus.

Microorganism

Kluyveromyces marxianus ZIM 1867 yeast strains were taken from the culture

collection of the Biotechnological Faculty, University of Ljubljana. The yeast strains stock

cultures were maintained on ATCC 200 solid slants medium at 30C.

Inoculum preparation

For the preparation of the working cultures, the yeast strains were re-inoculated into

flasks containing 50 mL of sterile liquid ATCC 200 and incubated aerobically under shaking

conditions (150 rpm) at 280C for 24 h. Sterile whey based media were inoculated with 5%

(v/v) of starter culture and then 5 mL 6 h old suspension was directly inoculated in 300 mL

Erlenmeyer flasks containing 50 mL medium.



28

Media composition

Composition of media based on whey used in laboratory experiments to obtain yeast

biomass is presented in Table 1. Sweet whey was supplemented with minerals and yeast

extract for an active multiplication of yeast cells.

Table 1 Different cultivation media for screening experiment

No. Sweet whey media

1 Non sterilized deproteinized* whey as a reference

2 Non sterilized deproteinized whey enriched with minerals: (NH4)2SO4, Na2HPO4,

MgSO4.7H2O, NaCl, CaSO4.2H2O, FeSO4.7H2O, ZnSO4.7H2O,

MnSO4.4H2O, CuSO4.5H2O, H3BO3

3 Non sterilized deproteinized whey enriched with (NH4)2SO4

4 Deproteinized and autoclaved** whey

5 Deproteinized and autoclaved whey enriched with minerals: (NH4)2SO4, Na2HPO4,

MgSO4.7H2O, NaCl, CaSO4.2H2O, FeSO4.7H2O, ZnSO4.7H2O,

MnSO4.4H2O, CuSO4.5H2O, H3BO3

6 Deproteinized and autoclaved whey enriched with (NH4)2SO4

7 Deproteinized and autoclaved whey enriched with yeast extract

8 Deproteinized and autoclaved whey enriched with (NH4)SO4 and KH2PO4

9 Deproteinized and autoclaved whey enriched with yeast extract, (NH4)SO4 and KH2PO4

10 Hydrolyzed and autoclaved whey

*Whey was deproteinized by thermal precipitation.

**Whey sterilization was performed by autoclaving at 1210C for 15 minutes.

Culture conditions

Batch fermentation was performed in 300 mL Erlenmeyer flasks, in aerobic conditions

(shaking at 150-200 rpm), pH=4-5, at 280C, for 24 h.

Analytical methods

At indicated time intervals, culture samples were collected for analysis of cell

concentration, cells viability and biomass estimation.

For cell counting a Thoma chamber with 20 µm depth was used. The yeast viability

was determined by staining yeast cells with methylene blue, which stains the dead yeast cells

which then appear as dark blue cells (data not shown).



29

Cell growth kinetics was determined on the basis of weight of dry biomass in

triplicate. A certain volume of the culture broth (5 mL) was centrifuged at 5500 rpm for 15

min. After two washing cycles with distilled water and centrifugations, the resulting pellets

were dried at 1050C, for 4 h.

Results and discussions

Composition of media is very important for the efficiency and reproducibility of

laboratory technology. According to Huang and Tang (2007), cultivation media should

contain all the necessary nutrients to maintain cell growth and to support product

biosynthesis. Preliminary laboratory experiments were performed comparatively using

synthetic and complex media, as a substrate for the development of the yeast strains. Initially,

K. marxianus cell growth was evaluated on non-deproteinized sterilized sweet whey and

deproteinized (permeate) sterilized sweet whey, respectively. The purpose of using non-

deproteinized sterile whey was economically to lower the cost of the process.

Utilization of whey proteins by K. marxianus is discussed controversially. Due to the

uncertainty of whey-protein assimilation by K. marxianus, it is assumed that whey proteins do

not contribute to assimilable nitrogen (Christian Löser, 2015), that is why the protein was

eliminated from whey.

After preparation of cheese sweet whey, inoculation of Kluyveromyces marxianus

ZIM 1867 yeast strains and incubation in suitable temperature, the biomass yield of the

culture of the isolated yeast strains was detected and is illustrated in figure 1.

Figure 1 Percentage variation of dry matter depending on the medium composition based on non-

sterilized deproteinized sweet whey



30

The types of whey used showed significant influence on the biomass production. For

complete consumption of lactose from whey, with accumulation of biomass, is necessary to

supplement the culture media.

When yeast strains were cultivated on non-sterilized deproteinized whey substrate,

which was prepared in several variances (Table 1), the highest amount of dry matter (1.78%)

was obtained in the sample of sweet whey enriched with minerals while whey medium

without any supplementation have given the lowest yield (1.51%). Amount of yeast biomass

production can be improved with addition of minerals or only ammonium sulfate as nitrogen

source. We found that ammonium sulfate (3g/L) had not significant effect on biomass yield

comparative with addition of minerals, but the amount of produced biomass increases from

1.51% in whey without supplementation to 1.67%, in the presence of nitrogen

supplementation. This is in agreement with results obtained by Hasani Ahmadreza

(2012),who obtained yeast biomass of Cryptococcus flavus and Candida curvata increased

from 15.68 and 10,17g/L in whey without supplementation to 17,12 and 12,34 g/L dry

biomass yield, respectively, in whey with nitrogen supplementation. The lowest cell biomass

recorded when whey permeate alone was used may be due to the fact that whey permeate lack

sufficient low molecular weight nitrogen, which presents a challenge to the growth of many

industrial microorganisms, so they often require supplementation (Adebayo-Tayo B, 2008)

High percentages of dry substance obtained compared to control (sweet whey) is due

to the fact that to yeast biomass added bacterial load existing in whey. On the other hand,

using these substrates is demonstrated that selected yeast strain is competitive with natural

micro-flora (lactic acid bacteria) present in unprocessed whey.

When sterilized deproteinized sweet whey was used as substrate in different types of

media compositions (Table 1) the highest percentages of dry matter (4.24%) was obtained on

whey supplemented with yeast extract, potassium dihydrogen phosphate and ammonium

sulfate (4.06%) (usual N supplement) and yeast extract alone (3.42%), respectively (figure 2).

Yeast extract is the most commonly used nitrogen source in fermentation as it

provides growth factors that are suitable for microbial growth (Hun 2013, Nancib N, 2001).

The yeast extract added in the culture medium, comes with the growth factors pantothenic

acid, inositol, thiamine, nicotinic acid, pyridoxine. The effectiveness of the yeast extract

supplementation for the production of Kluyveromyces yeast strain biomass has been studied

by Schultz N (2006) and Parrondo J (2009), who observed a stimulation of K. marxianus

growth, in sweet whey medium with yeast extract. As yeast extract supplementation is not



31

economically attractive, mixture of (NH4)2SO4, KH2PO4 and yeast extract was used and yeast

biomass production was higher (4.06% dry matter).

Figure 2 Percentage variation of dry matter depending on the medium composition based on sterilized

deproteinized sweet whey

To develop a microorganism both carbon and nitrogen source are essential. The effects

of distinct nitrogen sources have been also tested by several authors in order to achieve high

microbial mass production. Numerous studies have been performed to select the best nitrogen

source which can maximize cell growth of K. lactis. These sources include ammonium

chloride, ammonium nitrate, soybean meal, yeast extract and amino acids (Hun CH, 2013).

It is known that the presence of amino acids as the nitrogen source in the culture

medium results in a considerable growth and development of yeasts. Yeasts are able to use as

nitrogen source not only inorganic nitrogen but also the organic one, derived from amino

acids. Free amino nitrogen derived in situ through the hydrolysis of proteins by a protease

could substitute for the exogenous nitrogen source (Thomas KC, 1990). Therefore, culture

media composition based on sweet whey hydrolyzed were prepared and tested. Whey

hydrolysis was performed with protease from Bacillus licheniformis, at a temperature of 600C

and pH 7. Making a comparative analysis between hydrolyzed whey and whey supplemented

with yeast extract (Figure 3), it is found that the highest percentage of dry matter was obtained

on sweet whey hydrolyzed medium (5.06%). The proteins of complex cheese whey



32

hydrolyzed to allow for utilization by micro-organisms have been shown to improve the

biomass production compared with mineral medium (Maullu C, 1999).

Figure 3 Percentage variation of dry matter on hydrolyzed sweet whey and sweet whey supplemented with

yeast extract

The positive effect of amino acids on the growth and fermentation rates has previously

been noted by Thomas KC (1990, 1992) who observed that the effectiveness of an amino acid

as a source of nitrogen may be linked to the ability of the yeast to use the carbon skeleton

derived from that amino acid.

Conclusions

Culture media used in fermentations should contain all necessary elements for the

synthesis of both the cell biomass and metabolic products; moreover, they must be

economically profitable.

On the basis of these experiments hydrolyzed sweet whey medium used as substrate

for yeast growth proved the high cell biomass promoter. It is of considerable value if a safe

microorganism likes K. marxianus can utilize whey permeate lactose to generate profitable

quantities of biomass, useful by-product.

Acknowledgments

The financial support of project OLIGOLAC Eureka E!335/2013 by Executive Agency for

Higher Education, Research, Development and Innovation Funding is acknowledged.



33

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used as a biocatalyst in the sustainable production of ethyl acetate, Energy, Sustainability and Society 2015, 5:2,

DOI 10.1186/s13705-014-0028-2.

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biomass from cheese industry by-products, J Appl Microbiol, 1999, 86, 182-186.

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biotechnology. Edited by Demian AL, Solomon NA. Washington DC: ASM Press, 1986, 122-136.

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different nitrogen sources on the production of lactic acid from date juice by Lactobacillus casei subsp.

rhamnosus, Bioresource Technology, 2001, 78, 149-153.

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marxianus fermentation with lactose-added whey. Brazil J Chem Eng., 2009, 26, 445-456.

Schultz N., Chang L., Hauck A., Reuss M., Syldatk C., Microbial production of single-cell protein from

deproteinized whey concentrates. Appl Microbiol Biotechnol 2006, 69, 515-520.

Siso MIG., The biotechnological utilization of cheese whey: a review, BioresTechnol, 1996, 57, 1–11.

Smithers GW., Whey and whey proteins—from ‘gutter-to-gold’, Int Dairy J, 2008,18, 695–704.

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fermentation of wheat mash, Can. J. Microbiol. 1992, 38:626–634.



35

RESEARCH ON THE CHARACTERISTICS OF OBTAINING YOGURT

FROM COW’S MILK AND SOY MILK

Butoi Simona1, Butoi Laurențiu1, Tiţa Mihaela2, Prăvariu Ileana3

1 Terezianum Technical College of Food Industry Sibiu, 2 Faculty of Agricultural Sciences, Food Industry and Environment Protection, “Lucian Blaga” University of

Sibiu 3 Polisano Medical Clinic

ABSTRACT

Demand and consumption of soy milk products has considerably increased lately, because they represent one

of the most popular alternatives to cow milk, ideal for people who are lactose intolerant, with a high content

of essential nutrients including fibre, vitamins, minerals and antioxidants. One of the greatest qualities of soy

milk is that it can reduce cholesterol and protect cardiovascular health.

This paper aims to highlight and analyse soy milk yoghurt compared to cow's milk yogurt. Furthermore, the

present study asssesses the benefits of the consumption of soy milk yogurt.

KEYWORDS: milk, yogurt, soy, nutrients

INTRODUCTION

Soy milk, also called the soybean drink, is a beverage made from soy beans and has

approximately the same amount of protein as cow's milk, although, by contrast, it has a

different amino acid profile, less saturated fat and no cholesterol content compared to cow’s

milk. Soy milk unlike cow's milk does not contain lactose, which shows a good alternative for

people who are intolerant to lactose, while some people consume it for ethical or health

reasons.

Fortunately, lactose intolerance can be easily treated through diet foods that contain

lactose free food products that have no milk in their composition.

Yogurt has a Turkish origin, this term referring to slightly acidic fermented milk

products, with a mixed culture of lactic acid bacteria: Lactobacillus delbrueckii ssp.

bulgaricus, Streptococcus thermophilus ssp. salivarius, sometimes with additional sweeteners



36

such as fructose, glucose and sugar. Since the end of the 1970s, in the US, the product made

from soy milk has been called "soy milk yogurt" and "soy yogurt".

One of the opportunities of replacing cow's milk yogurt is to consume soy milk yogurt

that has a content of 0% lactose and is beneficial for people who are lactose intolerant.

Another benefit of soy yogurt consumption is the probiotics content. It is a well-

known fact that soy yogurt is produced by the same technology as the cow’s milk product,

both contain "healthy bacteria" such as probiotics, but during pasteurization some probiotic

cultures are destroyed, being reintroduced in yogurt after the pasteurization stage.

Unlike cow's milk yogurt, soy yogurt contains a significant amount of isoflavones and

is rich in omega-3 fatty acids, considered essential.

Soy yogurt is suitable for vegetarians, ovo-vegetarians, the Hebrew people, for people

with phenylketonuria or other dangerous diseases and for those with lactose intolerance or

allergy to milk.


As raw materials, the following were used: cow's milk from a private farm, soy milk,

starter cultures of lactic bacteria specific to yogurt.

The methods of analysis used were:

Water activity realized with moisture analyser Aqualab

Water activity is determined by the formula:

aw = or aw = x 100%;

where aw - water activity, expressed in%;

Determination of acidity – the Thorner method

Acidity (ºT) = V (ml of NaOH 0.1 N) x 10

Determination of dry matter content by using the ML - 50 thermobalance

Determination of protein titre – according to analytical standards in force

Calculation: protein titre = (V / 2) x f

in which:



37

V - volume NaOH 0,143n for the last titration;

f - factor of the NaOH solution 0,143n used in the last titration.

Determination of viscosity with the Brookfield YR-1 rheometer

Determination of lactose content – the polarimetric method II with automatic polarimeter.

RESULTS AND DISCUSSION

Water activity

The results obtained upon the analysis of determining water activity are set in the following

table:

Table 1. Water activity

Water activity in cow's milk yogurt

(%)

The water activity in soy milk

yogurt (%)

0,997 0,998

It is observed that yogurt made from soy milk has a water activity higher than cow's

milk yogurt, due to the dry matter content.

Acidity

The results upon determining the acidity are shown in the following table:

Table 2. Determination of acidity

Acidity of cow's milk yogurt (ºT) Acidity of soy milk yogurt (ºT)

88 146

Following the results in the table above, we observe that soy milk yogurt has a much higher

acidity than cow's milk yogurt, due to the chemical composition of soybeans.

Dry matter

The results upon determining the dry matter are presented in following table:

Table 3. Determination of dry matter

Dry matter in cow's milk yogurt (%)

Dry matter in soy milk yogurt (%)

11,5 10,3

It can be noted that the amount of dry substance in cow's milk yogurt is greater than the dry

substance in soy milk yogurt, due to higher dry matter content.



38

Protein titre

The results after determining protein titre are presented in following table:

Table 4. Determination of protein titre

Protein titre of cow's milk yogurt (%) Protein titre of soy milk yogurt (%)

0,2574 0,3575

After determining the protein titre, we could note that the value obtained from soy milk

yogurt was higher than that obtained from cow's milk yogurt, which shows that soy milk

yogurt has a higher percentage of protein content compared to cow's milk yogurt.

Viscosity

Upon determining the viscosity, the results presented in the following table were obtained:

Table 5. Determination of the viscosity

Viscosity Cow's milk yogurt

(mPa • s/cP)

Soy milk yogurt

(mPa • s/cP)

Start 1700 423,5

After 5 minutes

cP 177,3 640,1

S 72 72

% 40,5 34,5

RPM 60 60

According to the results from the table above, we observe that soy milk yogurt has a higher

viscosity than cow's milk yogurt, it is denser due to the chemical composition of soybeans.

Lactose

The results following the determination of lactose are presented in the table below:

Table 6. Determination of lactose

Determination of

lactose

Cow's milk yoghurt Soy milk yogurt

Rotation angle of

polarimeter

7,418 -

Lactose 7,06 -



39

It is noted that cow's milk yogurt has a lactose content of 7.06, as lactose is specific to dairy

products, not to vegetable products. The only source of lactose for human beings is milk.

CONCLUSIONS

Water activity is higher in soy yogurt milk than in cow's milk yogurt. The acidity is higher in

soy milk yogurt than in cow's milk yogurt. The dry substance percentage is higher in cow's

milk yogurt than in soy milk yogurt.

The soy milk yogurt has higher protein content than cow's milk yogurt.

The viscosity in soy milk yogurt is higher than in cow's milk yogurt.

The lactose content is only present in cow's milk yogurt.

REFERENCES 1. C.Banu, Camelia Vizireanu, „Procesarea Industrială a laptelui”, Editura „Tehnică”,București, 1998.

2. C.Banu, „Tratat de industrie alimentară-Tehnologii alimentare”, Editura ASAB, București, 2009.

3. Emil Rădulescu, „Alimentație inteligentă”, Editura „Viață și sănătate”, București, 2003.

4. George.Chintescu, Ștefan. Grigore ,, Îndrumător pentru tehnologia produselor lactate", Editura

Tehnică,București, 1982.

5. Gheorghe Menicinicopschi, „Si noi ce mai mȃncăm?”,Vol.I, Editura Coreus, București, 2010.

6. Pop.Flavia, ,,Îndrumător de laborator pentru tehnologia laptelui și a produselor lactate", Editura Risoprint,

2008

7. Tița Mihaela-Adriana, „Manual de analiză și controlul calității ȋn industria alimentară”, Editura

Universității „Lucian Blaga”, Sibiu, 2002.



40

IMPROVING GENETIC RESOURCES OF MICROORGANISM’S

VINEYARDS IN ORDER TO OBTAIN A HIGH QUALITY WINES,

VINEYARDS FROM LOCAL AREAS Diana Ionela Stegărușa, Ecaterina Lengyelb, Ovidiu Tițab, Ciucure Corina Teodoraa a National Institute for Research and Development for Cryogenics and Isotopic Technologies - ICIT Rm. Valcea,

4th Uzinei Street, PO Raureni, Box 7, Ramnicu Valcea, 240050, Valcea, Romania b Faculty of Agricultural Sciences, Food Industry and Environmental Protection/”Lucian Blaga” University of

Sibiu, 10th Victoriei Bd, Sibiu, 550024, Sibiu, Romania

Abstract The purpose of this paper is to identify and select a number of yeasts with biotechnological superior

properties to improve and enrich autochthonous genofund. This paper focuses on the isolation of a pure

culture of these yeasts and their identification using a rapid tests API 20C AUX. Monitoring alcoholic

fermentation was performed using laboratory fermenter system with a capacity of 2 L, equipped with sensors

for O2, CO2, pH, temperature, biomass, conductivity, results were captured on a data acquisition.

Obtained strains were multiplied in the culture media enriched with sucrose and ammonium sulphate in order

to amplify the biotechnological characteristics in order to obtain high quality wines, specific to local areas.

Isolated over 120 wine yeast strains, of which most have shown the ability to contribute to obtain wine with

superior aromatic and sensory qualities.

Keywords: yeast, API 20C AUX test, alcohol fermentation

Introduction

Wine Yeasts are essential in the technological processes of fermentation of grape must that

contribute to the award of the specific character wine area they come from (Radden, 2007). In

order to preserve these qualities are increasingly require the selection of native strains with

superior biotech characters so as to eliminate possible variety uniform taste assigned using

commercial yeast (Dubourdie et al. 2006, King et al. 2011, Pretorius 2009).

On the surface of grapes and vine is a wide range of microorganisms reached due to insects

and through earth and dust particles, precipitation, air currents which form the so-called

epiphytic microflora of grapes. The nature and number of these organisms are dependent on

several factors including the health and integrity of grapes and how plantation age, climatic



41

conditions. With the processing of grapes the microorganisms arive in the must, where,

depending on the temperature, composition of medium, the amount of sulfur dioxide, there is

a selection in the wort leaving only those microorganisms which are adapted better and was at

the begin in a larger number. Microbiota grape is rich in specific yeast species of the

plantation of origin of the variety, which is in direct correlation with the region's climate, soil

structure and other factors of " terroir ". In pressing phase in must are meet genres such as

Saccharomyces, Torulopsis, Hansenula, Pichia Kloeckera, Hanseniospora, Candida,

Brettanomyces associated with lactic bacteria, acetic, including Penicillium such as

Aspergillus, Geotrichum, Trichoderma, Absidia, Botrytis. Existing microorganisms must

resist more or less during alcoholic fermentation, many failed to adapt to environmental

factors created by these processes. Yeasts of the genus Saccharomyces presents an

exponential multiplication activity they managed to dominate the fermentation process,

eventually leading to the production of wines with higher alcohol concentrations (Eglinton et

al. 1996, King et al. 2008, Pretorius 2005b). These yeasts show the property to give specific

characters wine plantation, enhancing a wide range of aromas that lead to the creation of the

wine typicity of origin. In order to obtain quality wines are recommended by selected yeasts

with high biotechnological properties, but to keep their typicality and authenticity of local

strains are recommended from local microbiota (Reynolds et al. 2007, Swiegers et al. 2005,

Swiegers et al. 2007) .


- Grape must come from vineyards viticole Ciumbrud, Drăgășani, Jidvei, Recaș,

Segarcea

- 2 L volume fermenter equipped with:

- Sensors O2 , CO2 , pH , temperature , biomass , conductivity

- Double jacket , water circulation pump , thermostat bath

- the temperature of fermentation was set at 18⁰C

- period of harvest yeast : daily for 10 days starting on the third day of fermentation

- speed of palette fermenter was set at 200 rev / min

- API 20C AUX -Tests , Biomerieux , France, to identify yeast

- M1- MMA culture medium ( malt worth agar , Scharlau , Barcelona ) , YPG ( yeast

- peptone - glucose ) , 5% sucrose , 0.5 g/ L ammonium sulfate



42

- M2- MMA culture medium ( malt worth agar , Scharlau , Barcelona ) , YPG ( yeast

- peptone - glucose ) , 10% sucrose, 0.5g / L ammonium sulfate

- M3 - MMA culture medium ( malt wort agar , Scharlau , Barcelona ) , YPG ( yeast -

peptone - glucose ) , 15% sucrose, 0.5g / L ammonium sulfate

- blades, Keyance fluorescence microscope, Biozero

For monitoring alcoholic fermentation and identifying strains of Saccharomyces

cerevisiae, were introduced into the fermenter 2 L of fresh grape originated from vineyards,

production of 2013. The temperature of process was set at 18⁰C, cooling is performed using

water that was pumped the double jacket of the fermentation tank sump thermostat. Sensors of

fermenter were connected to a data acquisition, the results obtained were processed using the

software of the system .

Identification of yeasts was performed using rapid tests API 20C AUX. The API is based on

the property of assimilating yeasts or some sugars, resulting in the numeric code that leads to

their identification. Procedure requires placing a fresh yeast culture in a test tube containing 2

mL medium API NaCl 0.85 %, provided that the turbidity of the suspension does not exceed 2

Mc Farland. Of this suspension 100 μL and inoculated into 7 mL medium API C containing:

Ammonium sulfate 5 g, monopotassium phosphate 0.31 g, dipotassium phosphate 0.45 g,

Disodium phosphate 0.92 g, Sodium Chloride 0,1 g, calcium chloride 0.05 g, magnesium

sulfate 0.2 g, 0.005 g L - Histidine, L - Tryptofan 0.02 g L - Methionine 0.02 g, 0.5 g gelling

agent, Solution with vitamins 1 ml, trace elements solution 10 ml, 1000 ml deionized water.

Using a pipette, the wells are filled with the final suspension and incubated for 48-72 hours at

29⁰ C. Let plus positive wells (stained), and then the obtained values are added together for

each group of the substrate resulting in a code of 6 numbers inserted into the catalog for the

web API, indicating that the species identified .

In order to characterize the yeast culture inseminations were performed on culture media must

malt agar. Developed cultures were isolated yeast colonies that were studied to research

microscope Keyance wet method between slide and slide preparations. Selected strains were

multiplied in nutrient culture medium malt worth with added sucrose and ammonium sulfate

(M1, M2, M3 ) .

Results and discussion

In the fermentation and harvesting processes after the completion of successive samples could

be isolated and identified by API 20C AUX test, 125 strains of yeast Saccharomyces



43

cerevisiae, which have been denoted by the initial area of origin followed by numbers from 1

to 25. Subsequently plated on nutrient medium strains described above were then selected

based on cultural aspects, cell size. From a cultural standpoint these yeasts has a light colour

to cream with a glossy, bulging. Figure 1-5 shows a wine yeasts isolated from each selected.

Figure 1. Saccharomyces cerevisiae culture C16

Figure 3. Saccharomyces cerevisiae culture R10

Figure 2. Saccharomyces cerevisiae culture D15

Figure 4. Saccharomyces cerevisiae culture J21



44

Figure 5. Saccharomyces cerevisiae culture S15

The dimensions of the microscop (Table 1) were selected only those strains which showed

peaks . Thus remained in question following strains : C12, C16, C24, D13, D15, D19, J7,

J11, J21, R5, R10, R18, S14, S15, S25.

Table 1. The average amount of yeast strains isolated from local strains

Strain

Size (μm)

M1 M2 M3

C12 4,5x6,7 4,5x8,2 3,8x7,7

C16 4,3x6,5 4,6x8,2 3,9x7,2

C24 4,2x5,7 5,2x8,3 4,2x7,1

D13 5,2x7,3 5,5x8,4 3,8x7,2

D15 4,9x6,8 4,9x8,2 3,3x7,0

D19 4,3x6,7 4,5x8,1 4,1x7,2

J7 5,3x7,4 5,5x7,8 3,8x7,5

J11 4,6x6,9 4,7x8,0 3,7x7,6

J21 4,3x6,8 4,5x7,2 3,4x7,3

R5 4,6x6,8 4,7x8,4 4,2x8,3

R10 5,3x6,3 5,5x8,2 4,1x8,1

R18 4,2x6,9 4,5x7,4 3,6x7,2

S14 4,5x7,2 4,5x8,3 3,5x7,5

S15 4,3x7,5 4,5x7,9 3,8x6,6

S25 4,4x7,1 4,7x8,5 4,1x7,3



45

It is noted that the use of the M2 culture medium is preferably selected in increasing the size

of yeasts which are of higher value to the media M1 and M3. A greater amount of

carbohydrates may cause stress on the yeast cell so that their developer can be easily

inhibited. If M1 environment resulting values ranged between a minimum 4,2x5,7 and a

maximum of 5,3x7,5 μm for M2 culture medium were obtained minimum 4,5x7,2 μm and

maximum of 5,5x8,4 μm. If the culture medium M3 results ranged from between minimum

3,3x7,0 μm and maximum 4,2x8,3 μm .

Conclusions

Isolated strains may lead to the formation of a library of cultures derived strictly from local

strains to preserve varietal typicality and authenticity in our country. By isolating the lees

from local varieties and improve their qualities biotechnological optimization of media can

reach areas typical starter cultures of origin. The results lead to the recommendation to use

these strains in winemaking processes so as to eliminate the uniformity of taste due to the use

of imported strains established.

References 1. Dubourdie, D., Tominaga, T., Masneuf, I., Peyrot des Gachons, C., Murat, M.,L., 2006, The role of yeasts

in grape flavor develpoment during fermentation: the example of Sauvignon Blanc, American Journal of Enology and Viticulture, 57, 81-88

2. Eglinton, J.,M., Henschke, P., A., 1996, Saccharomyces cerevisiae strains AWRI 838, Lalvin EC1118 and Maurivin PDM do not produce excessive sulfur dioxide in white wine fermentations, Australian Journal of Grape and Wine Research, 2, (2), 77-83

3. King, E. S., Swiegers, J., Travis, B., Francis, I. L., Bastian, S. E., Pretorius, I. S. 2008, Coinoculated fermentations using Saccharomyces yeasts affect the volatile composition and sensory properties of Vitis vinifera I. cv. Sauvignon Blanc vines, Journal of Agricultural and Food Chemistry, 56(22), 10829-10837

4. King, E.S., Francis, I.L., Swiegers, J.H. and Curtin, C., 2011a Yeast strain-derived sensory differences retained in Sauvignon blanc wines after extended bottle storage. American Journal of Enology and Viticulture 62, 366–370

5. Pretorius I. S., 2009, Los nuevos retos en microbiologia del vino. Levaduras no productoras de SH₂, Seminario Téchnico Compuestos azufrados volatiles en vinos Problemas de reducción y aromas varietales, 18-29

6. Pretorius I.S. , 2005b, Modulation of volatile thiol and ester aromas in wine by modified wine yeast, Proceedings of the Weurman flavour research symposium, Roskilde, Denmark, 21-24 June 2005, Developments in Food Science, Elsevier, Amsterdam, The Netherlands

7. Radden, Rosemary, 2007, "Grapes and Wines of the World". The State Library of South Australia, GPO Box 419, Adelaide SA 5001. Retrieved, 06-16

8. Reynolds A. G., Schlosser J., Sorokowsky D., Roberts R., Willwerth J. and de Savigny Ch., 2007, Magnitude of Viticultural and Enological Effects. II. Relative Impacts of Cluster Thinning and Yeast Strain on Composition and Sensory Attributes of Chardonnay Musqué Am. J. Enol. Vitic., 58:25-41



46

9. Swiegers J.H., Bartowsky E.J., Henschke P.A. and Pretorius I.S., 2005, Yeast and bacterial modulation of wine aroma and flavor, Australian Journal of Grape and Wine Research, volume 11, Issue 2, 139-173

10. Swiegers, J.H. &I.S. Pretorius, 2007, Modulation of volatile sulfur compounds by wine yeast, Applied

Microbiology and Biotechnology 74:954-960



47

COMPARATIVE STUDY ON NICKEL REMOVAL FROM AQUEOUS

SOLUTIONS BY FLOTATION WITH ANIONIC AND CATIONIC

COLLECTOR

Adrian TURTUREANU

“Lucian Blaga” University of Sibiu, Romania, Faculty of Agricultural Sciences, Food Industries and

Environmental Protection

Abstract: The paper presents nickel removal by flotation from dilute aqueous solutions, at a laboratory scale, using an anionic collector (oleic acid) and a cationic collector (octadecylamine). The optimum values of the main parameters influencing this process were determined: pH of Ni(II) solutions, molar ratio collector : Ni(II), air flow rate, flotation time, initial concentration of Ni(II) and temperature. Using the optimal conditions, a high removal degree of nickel (> 98%) was obtained with both collectors.

Keywords: nickel, removal, flotation, anionic collector, oleic acid, cationic collector, octadecylamine

Introduction

Heavy metals are considered as hazardous pollutants due to their toxicity even at low

concentration, and non-biodegradable properties. The transportation and bioavailability of

heavy metals in the aquatic system is affected greatly by their binding to surfaces of solid

phases and complexion with the ligands in the water [1].

Heavy metals can cause accumulative poisoning, cancer, and brain damage when they

are found above the tolerance levels [2].

Exposure to nickel can cause dermatitis and allergic sensitization. About 10% of

women and 2% of men in the global population are highly sensitive to nickel. A part of these

sensitive people can develop skin rash called nickel dermatitis if they are exposed to nickel

through direct contact. This is the most common type of reaction to nickel exposure.

With nickel toxicity the type of nickel (speciation) is very important. Its form and the

ease with which it dissolves in water, affects the toxicity and the consequences for human

health. The most serious health effects occur when nickel is inhaled: increased risk of

respiratory cancer due to chronic inhalation of fumes or fine particles when exposure is to

known carcinogenic forms like nickel oxide; asthma, nasal and sinus problems [3].



48

In animal studies, which are much more extensive, nickel exposure caused: decreased

body weight, increase in still births, decrease in birth weight of offspring, and even death,

when nickel exposure is very high.

Nickel is necessary for a healthy plant growth. It is an important component of an

essential enzyme. At a level of 100 ppm or higher, nickel is considered to be phytotoxic,

harmful or toxic to plants [3].

The concentration of heavy metals in the industrial wastewater should be decreased to

the permissible limits. Chemical precipitation [4-8], coagulation [6], solvent extraction [9-

12], membrane separation [13-15], ion exchange [16, 17], adsorption [18-20] and foam

flotation [21-24 are commonly applied in the treatment of industrial effluents. Foam flotation

presents high selectivity and efficiency low space requirements, rapid operation, flexibility,

production of small volume of sludge and moderate costs [25].

The aim of this study is to compare the nickel removal efficiency from dilute aqueous

solutions by flotation, using anionic collector (oleic acid) and cationic collector

(octadecylamine).


Chemicals:

- nickel sulphate (NiSO47H2O p.a.) stock solution (2 g Ni(II)/dm3) from which

were prepared solution with 100 mg Ni(II)/dm3;

- anionic collector, oleic acid (named HOL), 0,25 M solution in ethanol;

- cationic collector, octadecylamine (named ODA), 0,25 M solution in ethanol;

- 15% and 1M NaOH solutions;

- 1M HNO3 solution.

Apparatus:

- bench-scale equipment for dispersed-air flotation technique;

- pH-meter WTW 96;

- atomic absorption spectrophotometer PYE UNICAM model SP 1900.

Working procedure:

The flotation bench-scale equipment consist of compressor, cock for air flow rate

adjustment, rotameter, 3,3 cm inner diameter glass flotation column (60 cm in height) with

porous glass frit (porosity G4).



49

From stock solution with 2 g Ni(II)/dm3 there were prepared solution with 100 mg

Ni(II)/dm3. The pH was adjusted to the desired value by adding NaOH or HNO3 and than it

was added the collector, oleic acid (HOL) or octadecylamine (ODA). The resulted solution

was submitted to flotation.

The main parameters influencing these processes were studied: pH of Ni(II) solutions,

molar ratio collector : Ni(II), air flow rate, time, temperature and initial concentration of

Ni(II) in sample. An atomic absorption spectrophotometer PYE UNICAM model SP 1900

was used to determine the nickel content of the solutions.

The nickel removal efficiency, R%, was calculated with the relation:

100)1(%0

CCR

where: C – concentration of Ni(II) after flotation;

C0 – initial concentration of Ni(II) in solutions.


The pH of the solution is one of the most important factors which influences the ion

separation by flotation, as it determines the magnitude and sign of the charge on the ions and

also the dissociation degree of the ionic groups of the surfactant molecules. Preliminary

experiments, shown in Figure 1, were conducted in order to determine the pH effect on the

nickel removal efficiency, (R%).

Figure 1. Influence of pH on the nickel removal efficiency Molar ratio collector : Ni(II)=1; air flow

rate=15 dm3/h, flotation time=15 min.

0

20

40

60

80

100

7 7,5 8 8,5 9 9,5 10 10,5 11 11,5 12

R %

pH

ODA

HOL



50

The two curves overlap almost perfectly. It can be seen that the flotation of nickel

ions, with both collectors, begins at pH = 8,5, but a good separation efficiency is obtained at

pH values greater than 9,5. At this pH values Ni(II) precipitates as hydroxide and flotation is

a precipitate flotation process.

The second studied factor was the effect of the molar ratio collector : Ni(II). As

surfactant was used oleic acid 0,25 M solution in ethanol and octadecylamine 0,25 M in

ethanol. Addition of ethanol as frother had the further advantage that the sizes of bubbles are

smaller, because of the lower surface tension of the solution. The results are shown in Figure

2. The increase of the molar ratio collector : Ni(II) determines a fast increase of removal

efficiency.

Figure 2. Effect of molar ratio collector : Ni(II) on the nickel removal efficiency

pH=10; air flow rate=15 dm3/h, flotation time=15 min.

The influence of air flow rate on Ni(II) removal efficiency is shown in Figure 3. The

maximum separation efficiency (over 98%) was obtained at low air flow rates. The increasing

of R% is faster with oleic acid than with octadecylamine. Increasing the air flow rate over 15

dm3/h, it can be seen a slow decrease of removal efficiency, but however not under 98%.

The influence of flotation time on Ni(II) removal efficiency is shown in Figure 4. It

can be seen that the separation process is very fast. In only 6 minutes is achieved a removal

efficiency of over 98%, for both collectors. However, the use of oleic acid is more effective

than octadecylamine.

60

70

80

90

100

0,25 0,5 0,75 1 1,25 1,5 1,75 2

R %

Molar ratio collector : Ni(II)

ODA

HOL



51

Figure 3. Effect of air flow rate on the nickel removal efficiency pH=10; molar ratio collector : Ni(II)=1,

flotation time=15 min.

Figure 4. Effect of flotation time on the nickel removal efficiency pH=10; molar ratio collector : Ni(II)=1, air

flow rate=10 dm3/h

9 1

9 2

9 3

9 4

9 5

9 6

9 7

9 8

9 9

1 0 0

3 6 9 1 2 1 5 1 8

R %

A ir flow rate , [dm 3 / h]

O D A

HO L

95

96

97

98

99

100

3 6 9 12 15 18

R %

Flotation time [min.]

ODA

HOL



52

Another factor influencing the flotation process is the initial concentration of Ni(II) in

solution. The results are shown in Figure 5. Decreasing Ni(II) concentration caused a marked

fall of removal efficiency, to 75%, for both collectors.

Figure 5. Effect of initial concentration of nickel on the removal efficiency. pH=10; molar ratio collector :

Ni(II)=1, air flow rate=10 dm3/h, flotation time=10 min.

Increasing temperature does not influence the removal efficiency with oleic acid, but

in case of octadecylamine the removal efficiency drops off dramatically, to 78%, (Figure 6).

60

70

80

90

100

100 80 60 40 20 0

R %

Initial concentration of Ni(II), [mg/dm3]

ODA

HOL

60

70

80

90

100

20 30 40 50 60

R %

Temperat ure [C ]

ODA

HOL



53

Figure 6. Effect of temperature on the nickel removal efficiency pH=10; molar ratio collector : Ni(II)=1, air

flow rate=10 dm3/h, flotation time=10 min.

Conclusions

The two studied collectors (oleic acid and octadecylamine) seem to have the same

behavior as collectors in nickel removal. Still using oleic acid leads to slightly better effective

separation. The use of octadecylamine is affected by temperature increase. High removal

efficiency (>98%) of Ni(II) from dilute aqueous solutions has been obtained by applying the

precipitate flotation process, at relatively shorter time than those used within other removal

methods.

A summary of the optimal conditions for nickel removal from synthetic solutions is

presented in Table 1.

Table 1. Optimum conditions for Ni(II) removal from synthetic solutions

Parameter Oleic acid (HOL) Octadecylamine (ODA) pH 10 – 12 10 – 12 Molar ratio collector : Ni(II) 1 – 2 1 – 1,75 Air flow rate, [dm3/h] 3 – 18 9 – 18 Flotation time [min.] 3 -18 6 – 18 Initial concentration of Ni(II) [mg Ni(II)/dm3]

80 – 100 80 – 100

Temperature [oC] 20 – 60 20 – 30

References:

1. Srivastava, V.C., Mall, I.D., Mishra, I.M., Modelling Individual and Competitive Adsorption of Nickel(II) and Zinc(II) Metal Ions from Aqueous Solution onto Bagasse Fly Ash, Sep. Sci. Technol., 2006, 41, 2685-2710.

2. Zouboulis, A.I., Matis, K.A., Lanara, B.G., Removal of nickel from dilute solutions by hydroxyapatite. II. Flotation studies, Sep. Sci. Technol., 1997, 32(10), 1755-1767.

3. http://www.incowatch.ca/content/4-2%20Toxicology.htm 4. Holtzman, S.A., Cyanide and Heavy Metal Removal: A Comparison of Different Chemistries with

Emphasis on an Innovative New Treatment Method, Sep. Sci. Technol., 1999, 34(14), 2761-2767. 5. Ennaassia, K., Kacemi, E., Kossir, A., Cote, G., Study of Removal of Cd(II) from Phosphoric Acid

Solutions by Precipitation of CdS with Na2S, Hydrometallurgy, 2002, 64(2), 101-109. 6. Charerntanyarak, L., Heavy Metals Removal by Chemical Coagulation and Precipitation, Wat. Sci.

Technol., 1999, 39(10-11), 135-141. 7. Youcai, Z., Stanforth, R., Selective Separation of Lead from Alkaline Zinc Solution by Sulfide

Precipitation, Sep. Sci. Technol., 2001, 36(11), 2561-2567.



54

8. Brooks, C.S., Recovery of Nonferrous Metals from Metal Finishing Industry Waste, Sep. Sci. Technol., 1993, 28(1-3), 579-587.

9. Mickler, W., Uhlemann, E., Liquid-Liquid Extraction of Copper from Ammoniacal Solution with 4-Acylpyrazol-5-ones, Sep. Sci. Technol., 1993, 28(10), 1913-1921.

10. Alguacil, F.J., Navarro, P., Non-Dispersive Solvent Extraction of Cu(II) by LIX 973N from Ammoniacal/Ammonium Carbonate Aqueous solutions, Hydrometallurgy, 2002, 65(1), 77-85.

11. Hu, S.Y.B., Wiencek, J.M., Copper-LIX 84 Extraction Equilibrium, Sep. Sci. Technol., 2000, 35(4), 469-479.

12. Lin, S.H., Huang, S.H., Juang, R.S., Nonideality in Two-Phase Systems of Copper(II) Extraction from Sulfate Solutions with LIX 64N in kerosene, Sep. Sci. Technol., 2002, 37(1), 147-155.

13. Ilias, S., Govind, R., A Study on Concentration Polarization in Ultrafiltration, Sep. Sci. Technol., 1993, 28(1-3), 361-369.

14. Mundkur, S.D., Waters, J.C., Polyelectrolite-Enhanced Ultrafiltration of Copper from a Waste Stream, Sep. Sci. Technol., 1993, 28(5), 1157-1163.

15. Orhan, G., Arslan, C., Bombach, H., Stelter, M., Nickel Recovery from the Rinse Waters of Plating Baths, Hydrometallurgy, 2002, 65(1), 1-9.

16. Yalcin, S., Apak, R., Hizal, J., Afar, H., Recovery of Copper (II) and Chromium (III, VI) from Electroplating-Industry Wastewater by Ion Exchange, Sep. Sci. Technol., 2001, 36(10), 2181-2187.

17. Karppnen, T.H., Pentti, A.Y., Evaluation of Selective Ion Exchange for Nickel and Cadmium Uptake from the Rinsewater of a Plating Shop, Sep. Sci. Technol., 2000, 35(10), 1619-1625.

18. Hail, C., Wales, D.S., Keane, M.A., Copper Removal from Aqueous Systems: Biosorption by Pseudomonas Syringae, Sep. Sci. Technol., 2001, 36(2), 223-229.

19. Bakkaloglu, I., Butter, T.J., Evison, L.M., Holland, F.S., Hancock, I.C., Screening of Various Types Biomass for Removal and Recovery of Heavy Metals (Zn, Cu, Ni) by Biosorption, Sedimentation and Desorption, Wat. Sci. Technol., 1998, 38(6), 269-277.

20. Sag, Y., Kaya, A., Kutsal, T., Biosorption of Lead(II), Nickel(II) and Copper(II) on Rhizopus arrhizus from Binary and Ternary Metal Mixtures, Sep. Sci. Technol., 2000, 35(16), 26-40

21. Edzwald, J.K., Principles and Applications of Dissolved Air Flotation, Wat. Sci. Technol., 1995, 31(3-4), 1-17.

22. Zouboulis, A.I., Matis, K.A., Removal of Cadmium from Dilute Solutions by Flotation, Wat. Sci. Technol., 1995, 31(3-4), 315-323.

23. Huang, S.D., Su, P.G., Huang, S.P., Ho, I.L., Tsai, T.Y., Adsorbing Colloid Flotation with Polyaluminum Chloride: A Powerful Technique for Removing Heavy Metals from Wastewater, Sep. Sci. Technol., 2000, 35(8), 1223-1231.

24. Zouboulis, A.I., Lazaridis, N.K., Zamboulis, D., Adsorption of Cr(VI) on Activated Carbon and Its Subsequent Separation through Foam Flotation, Sep. Sci. Technol., 1994, 29, 385-395.

25. Matis, K.A., Zouboulis, A.I., Flotation techniques in water technology for metals recovery, Sep. Sci. Technol., 2001, 36(16), 3777-3800.



55

NATURAL ANTIOXIDANT FEATURE ESTIMATED IN NON-

ALCOHOLIC DRINKS BY CHEMILUMINOMETRY

DARIE Neli

Faculty of Agricultural Sciences, Food Industry and Environmental Protection,“Lucian Blaga” University of

Sibiu, Romania, [email protected]

Abstract

Natural antioxidant feature from non-alcoholic drinks as sea buckthorn hydrolytic extract, rosy grapefruit

juice, kiwi juice, rose hip hydrolytic extract, red berries juice and goji berries juice were estimated by

chemiluminometry. The antioxidant capacity protecting from the free radicals was measured in

forementioned drinks, as ascorbic acid units.

Keywords: natural antioxidant features, chemiluminometry, free radicals, non-alcoholic drinks

INTRODUCTION

Some natural non-alcoholic drinks ordinary used in the human diet could be interesting

by their content in antioxidants [5]. They have total antioxidant capacity (TAC) and a

hydrolytic phase antioxidant capacity (HPAC). The body deseases caused by oxidative stress

and induced by the free radicals [1, 8, 4] could be prevent by different food antioxidants as

polyphenols, vitamins (C, E), flavonoids, carotenoids etc. Though the free radicals have a

short life in the biologic cells, they modify proteins and also unsaturated fatty acids (in the

lipid membranes). The antioxidative natural systems developed in cells protect aginst damage

through these radicals [7, 4].

In present study, the chemiluminescence assay [2,3,6] has been applied to evaluate a

total antioxidant capacity and a hydrophilic phase antioxidant capacity comparatively in

freshen and coolness keeping of six non-alcoholic drinks rich in antioxidants, that could be

important from the nutritional point of view, in order to establish the main contributors to the

TAC of healthy antioxidative diets. Also, drinks with high TAC values were chosen and

characterized for antioxidants natural sources cocktail.



56


The six evaluated drinks samples were: sea buckthorn (Hippophaë rhamnoides L.)

hydrolytic extract, rosy grapefruit (Citrus paradisi) juice, kiwi (Actinidia deliciosa) juice,

rose hip (Rosa canina) hydrolytic extract, red berries (Ribes rubrum) juice and goji berries

(Lycium barbarum Ningxia) juice. The drinks samples were diluted aliquots in water, then

mixted with etyl acetate (in 1:1 ratio) and shaking for 20 min. After centrifugation at 3000

g/5 min., the antioxidant capacity with hydrophilic and lipophilic compounds separately was

measured in the aqueous phases, using volumes of 10; 20; 30; 40 and 50 μL of samples.

For the experimental researches there were employed as reagents photo-sensitizer for

water-soluble/fat-soluble substances and detection reagent (Analytik Jena AG), buffers,

sample solvent, acid ascorbic as calibration standard to cuantify the water and fat soluble

antioxidants (in equivalents of ascorbic acid). The total antioxidative capacity (TAC) of

antioxidants in the six drinks studied was measured following the increasing of the

chemiluminescence, in a photochemiluminometer (method PCL as described in Analytik Jena

assay with a Photochem device).


One the purpose of this study was to measure and compare the antioxidant capacities

of six customary non-alcoholic drinks as: sea buckthorn hydrolytic extract, rosy grapefruit

juice, kiwi juice, rose hip hydrolytic extract, red berries juice and goji berries juice. The

maximum measuring times was about 2 minutes. The measurements were done in aquous

phase and blank values are measured after 40 seconds.

Fig. 1 shows the results for total antioxidant capacity (TAC), expressed as ascorbic

acid units/ml drink sample after instant dressing. The different values of TAC are in agree

with the drink chemical antioxidant composition. Experimental data have shown a high

chemiluminescence for non-alcoholic drink, in rose hip (extract), sea buckthorn extract and

goji berries juice, cause of rich phenolic constituents and a high TAC. The rose hip extract,

sea buckthorn hydrolytic extract and goji berries juice having the greatest TAC value were

used therefore for an antioxidant cocktail non-alcoholic drink and tested in hydrophilic

fraction. The freshen of rose hip, sea buckthorn and goji, with verified antioxidant features



57

and high experimental values of TAC were preserved at 4 degrees Celsius, in order to follow

some possible changes in antioxidant capacity.

050

100150200250300350400450

kiwi juice rose hiphydroliticextract

seabuckthornhydroliticextract

rosygrapefruit

juice

red berriesjuice

goji berriesjuice

1 2 3 4 5 6

TAC (ascorbic acid units/ml sample)

Figure 1. TAC data for some natural non-alcoholic samples

Fig. 2 and table 1 indicates the evolution of TAC values after times preservation

without be chemical preserved. TAC was measured after 24 hours, 48 hours, 4 days and 7

days of coolness keeping. As to obtain a high and fresh TAC cocktail, the samples having

highest TAC (of rose hip, sea buckthorn and goji berries) were kept. They have sensibly lost

antioxidant capacity even till 51%, after 7 days of the coolness keeping.

0

50

100

150

200

250

300

350

400

450

24 h 48 h 4 days 7 days

TAC (ascorbic acid units/ml sample)

1 rose hip hydrolit ic extract

2 sea buckthorn hydrolit ic extract

3 goji berries juice

Figure 2. Antioxidant capacity changes after times preservation



58

Table 1. Samples TAC decreasing in coolness

keeping.

Samples

TAC evolution in coolness keeping (%)x

time coolness keeping (days)

2 4 7

rose hip hydrolytic extract 1,24 9,43 45,41

sea buckthorn hydrolytic extract 5,17 16,79 51,93

goji berries juice 9,83 17,75 45,62 x reproductibility CV 1,8%

The hydrophilic samples phase has major contribution in TAC. Table 2 presents total

antioxidant capacity values of the studied non-alcoholic samples in their HPAC. High values

have also found in hydrolytic phase of rose hip, sea buckthorn and goji berries juices and the

data prescribe these as valuable compounds of an antioxidative cocktail.

Table 2. TAC samples joined at the hydrophilic phase.

Samples HPAC (as acid ascorbic units, mmol/l)x

goji berries juice 295,65

sea buckthorn extract 326,33

rose hip extract 347,25

CONCLUSIONS

The TAC and HPAC, measured by chemiluminescence, in six non-alcoholic drinks, in

freshen and coolness keeping prove three major antioxidative sources valuable for healthy

diets (rose hip, sea buckthorn and goji berries juices).

REFERENCES

1. Brighenti, F., Valtuena, S., Pellegrini, N., Ardigo, D., Del Rio, D., Salvatore, S., Total antioxidant capacity of

the diet is inversely and independently related to plasma concentrations of high-sensitive C-reactive protein in

adult Italian subjects. British J. Nutrition , 93(5), 2005, p. 616-625

2. Buchring, M., Popov, I., Lewin, G., Stange , R., Phys. Chem. Biol. & Med., 1994, 96, p.100



59

3. Halliwell, B., Gutteidge, J.M.C., Arch. Biochem. Biophys., 1990, 280, p.1-8

4. Goetzke B., Nitzko S., Spiller A., Consumption of organic and functional food. A matter of well-being and

health?, Appetite 77C, 2014, p. 94–103

5. Lugasi, A., Hovari, J. Antioxidant properties of commercial alcoholic and nonalcoholic beverages.

Nahrung/Food, 47(2), 79-86, 2003

6. Popov. I., Lewin, G., Methods in Enzymology, vol.300, 1999, p. 437-456

7. Robinson, E.E., Maxwell, S.R.J., Thoerpe, G.H.G., An investigationof the antioxidant activity of black tea

using enhanced luminescence, Free Radical Research, i1997, 26, p.291-302

8. Serafini, M., Laranjinha, J.A., Almeida, L.M., Maiani, G., Inhibition of human LDH lipid peroxidation by

phenol-rich beverages and their impact on plasma total antioxidant capacity in humans. Journal of Nutritional

Biochemistry, 2000, 11, p.585-590.



60

RESEARCHE REGARDING ANODIC OXIDATION OF ALUMINIUM

FROM ACID MIXTURE ELECTROLYTE

Carmen Monica Cretu

Lucian Blaga” University of Sibiu, Faculty of Agricultural Sciences, Food Industry and Environment

Protection, 7-9 Dr. Ion Raţiu St., 550012 Sibiu, Romania

Abstract The paper presents the study of anodic oxidation of aluminium using a sulphuric and oxalic acid mixture

electrolyte. The influences of current density, acid mixture concentration and temperature on current

efficiency and both the thickness and aspect of the deposited Al2O3 were studied. Optimum sulphuric and

oxalic acid concentration, current density and temperature values were determined.

Keywords: anodic oxidation, aluminium, sulphuric acid, oxalic acid, Al2O3, current efficiency, thickness

Introduction

The electrodeposition of Al2O3 layer on the aluminium surface and its alloys presents

importance for the economic process. The method is known in our country with the eloxation

term for the anodic oxidation of aluminium.

The employed acid electrolytes for anodic oxidation of aluminium are based on

sulphuric acid [1-4], oxalic acid [5, 6], phosphoric acid [7, 8], chromic acid [9,10] and acid

mixtures [11-14].

The study presents the influence of production parameters on anodic oxidation of

aluminium in mixture of sulphuric and oxalic acid electrolyte. The current efficiency and both

the thickness and aspect of the deposited Al2O3 were determined.

Materials and Methods

The reagents used were of analytical grade: H2SO4 96% p.a., oxalic acid p.a., NaOH

p.a. and HCl 36.5% p.a. All solutions were prepared with double distilled water.

The mixture acid electrolyte used for this study of anodic oxidation of aluminium had

the concentration between 0,1 - 0,5 M.



61

Before the electrodeposition experiments, the aluminium electrodes were polished with

abrasive paper and then submitted to chemical degreasing in an aqueous solution of sodium

hydroxide 20% and activation in HCl 1:1 solution.

The anodic oxidation of aluminium was performed by galvanostatic electrolysis, in

different experimental conditions for 15 minutes, in an Hull stiplex cell, with a capacity of 247

cm3. The anode was an aluminium strip, with 99.9% purity. The cathode was a lead strip. A

Princeton Applied Research galvanostat, model 175, [4] was used for the current density.

The thickness of the deposited Al2O3 on aluminium anode was determined by a

metallographic microscope MMB 2500. The aluminium anode was weighed on an analytical

balance before and after each anodic oxidation.

After anodic oxidation of aluminium, the anode was immersed for 5 minutes in a

double distilled water at 90 oC, in order to improve the corrosion resistance and to decrease the

porosity of Al2O3 layer.


The protective properties of Al2O3 layer depend from the nature and the electrolyte

concentration and the production parameters.

The effect of current density

The anodic oxidation of aluminium was studied at 25 oC, using a mixture of the same

volume of 0,4 M sulphuric acid and 0,3 M oxalic acid solution. The effect of current density on

current efficiency and both the thickness and aspect of the deposited Al2O3 was studied.

The current densities employed were between 1 and 2,5 A dm-2.

The results concerning the effect of current density on current efficiency of anodic

oxidation of aluminium and on Al2O3 layer thickness and aspect are presented in Table 1.

Table1 The effect of current density on current efficiency and on Al2O3 deposit thickness and aspect: 25 oC;

sulphuric acid concentration = 0,4 M; oxalic acid concentration = 0,3 M.

current density

(A dm-2)

current efficiency

(%)

Al2O3 thickness

(m)

Al2O3 deposit

aspect

1 82 1,12 dull, porous



62

1.5 88 1,28 bright

2 92 1,36 bright

2,5 86 1,18 pulverous

The experimental results show an increase and then a decrease of current efficiency when the current density increases.

At low current densities, the Al2O3 film is dull and porous, while at very high current

densities, it becomes pulverous because there is an heating of mixture acid solution by Joule

effect which produces a dissolution of Al2O3 deposit.

The optimum current density is 2 A dm-2 and in this case, the Al2O3 layer is bright,

adherent and continuous and the current efficiency and the thickness of film are maximum.

The effect of mixture of sulphuric acid and oxalic acid concentration

The influence of mixture of sulphuric acid and oxalic acid concentration on current

efficiency and both the thickness and aspect of the deposited Al2O3 was studied at a current

density of 2 A dm-2 and at 25 oC.

The mixture acid concentration was varied from 0,1 to 0,5 M.

The results concerning the effect of mixture acid concentration on current efficiency

and on the thickness and aspect of the deposited Al2O3 layer are presented in Table 2.

Table 2 The effect of mixture of sulphuric acid and oxalic acid concentration on current efficiency and on Al2O3

deposit thickness and aspect: 25 oC, i = 2 A dm-2.

sulphuric acid

concentration

(M)

oxalic acid

concentration

(M)

current

efficiency

(%)

Al2O3

thickness

(m)

Al2O3 deposit

aspect

0,2 0,1 81 1,14 dull, porous

0,3 0,2 86 1,28 bright

0,4 0,3 90 1,44 bright

0,5 0,4 94 1,56 pulverous

The increase of mixture of sulphuric acid and oxalic acid concentration leads to an

increase of current efficiency of anodic oxidation of aluminium. Increasing the mixture of

sulphuric acid and oxalic acid concentration results in an increase of Al2O3 film thickness.



63

At 0,2 M sulphuric acid concentration and 0,3 M oxalic acid concentration, the Al2O3

layer is dull and porous.

The 0,5 M sulphuric acid concentration and 0,4 M oxalic acid concentration is not

advisable for anodic oxidation of aluminium because leads in the producing of pulverous

Al2O3 deposit which is not for decorativ aspect. In this case, the dissolution rate of Al2O3 film

is greater than its forming rate and result pulverous deposits.

The values of current efficiency show an electrolyte concentration of 0,3 - 0,4 M

sulphuric acid and 0,2 – 0,3 M oxalic acid for the producing of bright, compact, adherent and

resistant Al2O3 deposits.

The effect of temperature

The anodic oxidation of aluminium was performed at a current density of 2 A dm-2,

using sulphuric acid concentration 0,4 M and oxalic acid concentration 0,3 M.

The tested temperatures were: 20 C, 25 C, 30 C and 35 C.

The effect of temperature on current efficiency and on the thickness and aspect of the

deposited Al2O3 layer is presented in Table 3.

Table 3 The effect of temperature on current efficiency and on Al2O3 deposit thickness and aspect:

i = 2 A dm-2, sulphuric acid concentration = 0,4 M; oxalic acid concentration = 0,3 M. temperature

( 0C )

current

efficiency

(%)

Al2O3

thickness

(m)

Al2O3 deposit

aspect

20 86 1,22 dull, porous

25 90 1,34 bright

30 94 1,48 pulverous

35 96 1,56 pulverous

The results show the increase of current efficiency with the temperature.

The Al2O3 thickness layer increases when the temperature increases.

At 20 0C temperature, a dull and porous Al2O3 deposit is obtained which is not good for

following protection. The optimum temperature for the anodic oxidation of aluminium is 25ºC

and the Al2O3 film is bright, continuous and compact. The dissolution process of Al2O3 deposit is

intensified by the increase of temperature, so that temperatures above 30ºC generate pulverous

Al2O3 coatings.



64

Conclusions

The goal of this set of electrodeposition experiments was to determine the optimum

conditions for the producing of bright, continuous, adherent and compact Al2O3 layers, at

high current efficiency.

The analysis of the experimental results can define preferred or optimum production

parameters, i.e. the conditions that produce good quality Al2O3 deposition, with anticorrosive

properties and pleasant aspect.

The conclusions of the study for anodic oxidation of aluminium from mixture of

sulphuric acid and oxalic acid electrolyte show that:

- The optimum current density is 2 A dm-2.

- The optimum sulphuric acid concentration is 0,4 M and oxalic acid concentration is 0,3

M.

- The optimum temperature is 25 C.

References 1. Holzer F., Muller S., Desilvestro J., Haas O., Aluminium alloys in sulphuric acid. Electrochemical

behaviour of rotating and stationary disc electrodes, J. Appl. Electrochem., 1993, 23(2), 125-134.

2. Graeve I.De, Terryn H., Thompson G.E., Influence of heat transfer on anodic oxidation of aluminium, J.

Appl. Electrochem., 2002, 32(1), 73-83.

3. Zhu X.F., Li D.D., Song Y., Xiao Y.H., The study on oxygen bubbles of anodic alumina based on high

purity aluminium, Mat. Lett., 2005, 599(24-25), 3160-3163.

4. Cretu, C.M., The Study of Aluminium Eloxation From Sulphuric Acid Electrolyte, Proceedings of the 7th

International Conference “Integrated Systems for Agri-Food Production SIPA’11”, Nyiregyhaza

(Hungary), 10-12 november 2011, 163-165, ISBN 978-606-569-312-8, 978-615-5097-26-3,

http://www.sipa11.hu.

5. Huang Q., Lye W.K., Reed M.L., Mechanism of isolated pore formation in anodic alumina, Nanotech.,

2007, 18(10), 452-458.

6. Li Z.J., Huang K.L., Electrochemical fabrication of sandwich nanostructures based on anodic alumina, J.

Braz. Chem. Soc., 2007, 18(2), 406-409.

7. Dasquet J.P., Bonino J.P., Caillard D., Bes R.S., Zinc impregnation of the anodic oxidation layer of 1050

and 2024 aluminium alloys, J. Appl. Electrochem., 2000, 30(7), 845-853.

8. Zuo Y., Zhao Y., Li X., Li N., Bai X., Qin S., Yu W., Synthesis of alumina nanowires and nanorods by

anodic oxidation method, Mat. Lett., 2006, 60(24), 2937-2940.

9. Stojadinovic S., Belca I., Zekovic L., Kasalica B., Nikolic D., Galvanoluminescence of porous oxide

films formed by anodization of aluminium in chromic acid solution, Electrochem. Communic., 2004,



65

6(10), 1016-1020.

10. Garcia Vergara S.J., Skeldon P., Thompson G.E., Habazaki H., A tracer investigation of chromic acid

anodizing of aluminium, Surface and Interface analysis, 2007, 39(11), 860-864.

11. Dasquet J.P., Caillard P., Bonino J.P., Bes R.S., Characterization of the protective effect of aluminium

surface treatments by d.c. and a.c. measurements, J. Mat. Science, 2001, 36(14), 3549-3555.

12. Shingubara S., Morimoto K., Sakaue H., Takahagi T, Self organization of a porous nanohole array using

a sulphuric/oxalic acid mixtures electrolyte, Electrochem. Solid-State Lett., 2004, 7(3), 15-17.

13. Guo C., Zuo Y., Zhao Z., Zhao X., Xiong J., Anodic oxidation parameters on Al-Cu alloy anodizing Film’s

electrochemical ability by EIS, Advanced Mat. Res., 2006, 11-12, 665-668.

14. Keshavarz A., Parang Z., Nasseri A., The effect of sulfuric acid, oxalic acid and their combination on the size and

regularity of the porous alumina by anodization, J. Nanostr. Chem., 2013, 3(34), 1-4.



66

BIOACTIVE PLANTS - VALUABLE SOURCE OF TANNINS

Daniela Maria ŞANDRU

PhD Fellow, SOP HRD/159/1.5/S/133675 Project, Romanian Academy Iasi Branch or “Lucian Blaga” University from Sibiu, Partner, e-mail: [email protected]

Abstract This study aims at highlighting the bioactive potential of some plants which are important sources of tannins.

In order to measure the concentration of tannins in the plants under experimentation, we employed the

method of tannin dosing using Potassium permanganate; as indicator, we used indigo carmine. The results

of this study show reveal that Vaccinium myrtillus is an important source of tannins, which is why it can be

used to obtain certain preparations.

Keywords: tannins, dosing, bioactive compounds, indigo carmine

Introduction

Tannins are polyphenolic organic substances whose molecular mass ranges between 500 and

5000 atoms. They have an astringent taste and, together with some proteins, they make up

impermeable and rot-proof compounds, a process underlying hide tanning. The term “tannin”

comes from technology and not from any chemical classification. Tannins are widely spread

in the plant kingdom [1].

The plants used in this experiment are extremely important, due to their composition and

their concentration of biologically active compounds. Cichorium intybus is very valuable and

is used as animal feed. People try to maintain this plant as animal fodder and also use it in

medicinal preparations [2]. Cichorium intybus was rather poorly studied, both in chemistry

and in pharmaceuticals. Over 100 compounds were isolated and identified in this plant, most

of them in the root. Most measurements were made on aqueous or alcoholic extracts [3].

Plant biomass can be affected by harsh environmental conditions associated with high

altitudes. The biomass of Achillea millefolium drops with the altitude. The flowers are richer

in substances that can be turned into antiallergenic compounds using steam. The dark blue

essential oil extracted through the steam distillation of flowers is generally used as an anti-

inflammatory in the treatment of colds. Besides, the essential oil is used as massage oil for

swollen joints [4].



67

Vaccinium myrtillus is a rich source of polyphenolic compounds and tannins implicitly.

Polyphenolic compounds confer blueberries their blue or black color and their high

concentration of antioxidants. Blueberries are efficient in the treatment of swellings,

dyslipidemia, hyperglycaemia, cancer, diabetes and other diseases. Blueberries are renowned

for their anti-cancer potential and some studies used blueberries, both alone and combined

with other fruits, to prove their action [5].

Foeniculum vulgare is a herb used both in medicine and in cuisine. It tastes like anise and is

one of the ingredients of absinthe. Fennel seeds taste and look like anise seeds, but they are

smaller. Fennel is used to flavor some natural toothpastes. Fennel seeds are used to make

delicious desserts [6].

Xanthium spinosum is a medicinal herb which can be found worldwide. Many of its

pharmaceutical properties are efficient in the treatment of certain conditions, such as:

diarrhea, swellings, snake bites, fever and liver conditions that are at least partially correlated

with the uncontrolled release of certain metabolites and arachidonic acid. In vitro ziniolide is

substance that inhibits phorbol, i.e. the key regulator of arachidonic acid.


Alcoholic extracts of plants under study: chicory, yarrow, blueberry, fennel, Xanthium

spinosum.

KMnO4 0.1N

Indigo carmine 0.1%

Concentrated sulfuric acid

We used the following method:

25 ml of the tannins extract is diluted with 200 ml distilled water, to which we add 25 ml

indigo carmine solution. The mix is agitated and titrated with the KMnO4 0.1N solution, until

the indicator turns from blue to gold-yellow, through blue-green. Meanwhile, we make a

blank sample only containing distilled water (225 ml) and indigo carmine (25 ml) titrated

with KMnO4.



68


Figure 1. The amounts of tannins calculated for the plant macerates under study

Figure 1 shows the comparative amount of tannins in the five plants in this experiment. We

can notice that the amount of tannins in Vaccinium myrtillus is aproximately 50% higher than

in Cichorium intybus . In the other extracts, the amount of tannins is much lower than in the

first two. It is noticeable that Achillea millefolium and Xanthium spinosum have an almost

equal concentration of tannins.

Conclusions

The results show that all extracts contain tannins. As expected, Vaccinium myrtillus has the

highest concentration of tannins, while Cichorium intybus has a medium amount of tannins.

The lowest values were recorded in Achillea millefolium, Xanthium spinosum and

Foeniculum vulgare.



69

Comparing our results to the results of other studies, we can state that the bioactive potential

of these plants is very high, which means that these extracts can be used in various

preparations which are welcome in traditional medicine.

ACKNOWLEDGEMENTS This paper is supported by the Sectoral Operational Programme Human Resources

Development (SOP HRD), financed from the European Social Fund and by the Romanian

Government under the contract number POSDRU/159/1.5/S/133675.

References 1. Istudor V. - Pharmacognosy, Phytochemistry, Phytotherapy, vol I, Medical Publishing House,

Bucharest, 1998, vol II (2001).

2. Ziamajidi N, Khaghania S, Hassanzadeh G, et al. Amelioration by chicory seed extract of diabetes- and

oleic acid-induced non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) via

modulation of PPAR-alpha and SREBP-1. Food and Chemical Toxicology. 2013;58:198–209.

3. Suresh B, Sherkhane PD, Kale S, Eapen S, Ravishankar GA. Uptake and degradation of DDT by hairy

root cultures of Cichorium intybus and Brassica juncea. Chemosphere. 2005;61(9):1288–1292.

4. Wood, John (2006). Hardy Perennials and Old Fashioned Flower. Project Gutenberg Literary Archive

Foundation.

5. Seeram N.P. Berry fruits for cancer prevention: Current status and future prospects. J Agric Food

Chem.2009;56:630–5.

6. Classification for Kingdo Plantae Down to Genus Foeniculum Mill. US Department of Agriculture,

Natural Resources Conservation Service. 2015. Retrieved 24 March 2015.

7. Biesalski H.K., Poliphenols and inflammation basic interactions. Curr. Opin. Clin. Nutr. Metab. Care

2007 10: 724-8.



70

USE OF LOCAL NATURAL RESOURCES AT IMPROVE THE

QUALITY PREMIXES PRODUCT FOR GFD Short communication

*Iancu Maria Lidia, **Iancu Bianca Maria *”Lucian Blaga” University of Sibiu, Faculty of Agricultural Science, Food Industry and Environmental

Protection Department of Agricultural Science and Food Industry, 5-7, Dr. Ion Ratiu str. România

tel.+04(0)269211338; *Corresponding author: [email protected] **"Lucian Blaga" University of Sibiu, Faculty of Medicine, 2A, Lucian Blaga Street, Sibiu,550169, România

Abstract

GFD(Gluten-free diet)is the only solution in solving problems given by celiac disease. Although there mixes

on the market and studies on the occasion this study I want to communicate that sees a cheaper and

friendlier as an alternative to existing ones. It was found that the potato pulp cooked, crushed added 35% to

the basic formula dough for gluten-free products can improve texture, specific volume (from 2.01 to 2,39 cm3·1g-1), elasticity (from 88 to 90 %) and taste characteristics of bread for GFD.

Keywords: gluten-free bread, non gluten ingredient, quality descriptive, potato

Introduction

Hence, a requirement to produce high quality and wide variety of gluten-free products

is as important as ever.

Gluten-free diet is proposed as a treatment of several illnesses, from which celiac

disease (CD) is the most significant, as the restrictive menu without gluten is the only

effective therapy for this affliation (El-Chammas K., et al, 2011).

Gluten was also found in a significant number of foods with gluten/wheat-related advisory

warnings (Sharma M.G., et al., 2015).

Different materials are used in the preparation of gluten-free products: vegetable flours

(yellow pepper, green pepper, red pepper, tomato, spinach, pumpkin, zucchini, carrot,

asparagus, fennel, egg an plant flours) (Padalino L., et al, 2013).

We use various materials that improve the nutritional and organoleptic characteristics of

gluten-free products such as, plant oil, gum guar, potato starch, corn starch, salt, sugar

(Alvarez-Jubete L., et al., 2010; Korus J., et al., 2015).



71

The migration of water from core to crust cause considerable damage to the quality of bread

during storage(Matos Segura and Rosel, 2011; 2015; O, Shea N., et al., 2014).

Currently utilized gluten-free flours include maize, potato and rice flour and starches, used as

base flours due to their bland flavor and neutral effects on baked products. These flours and

starches usually tend to be low in nutrition and have very minimal structure-building

potential.

Because the market in Romania, especially in Transylvania are not always mixes on the

market, and ingredients are only in specialist shops is proposed study. Were tested a few

ingredients, natural, like milk and egg. For raising used as an alternative to other studies

baking powder. Finally aimed to influence the macroscopic characteristics of gluten-free

products, of boiled potato added in a proportion of 35%. Has been replaced with potato pulp

35% a mixture of ingredients. This mixture was obtained by RSM and published (Iancu ML.,

et al., 2015). The reason was to try to improve in particular taste characteristics and keeping

time freshness.

Materials and method

Recipe manufacturing, operational parameters and analysis methods of bread : To prepare

the dough was done: for control sample (Gl-Fr1) corn starch, yeast, salt, xanthan gum and

water in a ratio of 1:15 to % dry matter (dry matter) of the starch. Was added to the basic

formula 1 egg (Gl-Fr2 ), less water needed for consistency as clay (Gl-Fr3); yeast 1,75% at

dough with egg (Gl-Fr4); 1 egg, raising agent: sodium bicarbonate instead of the yeast(Gl-

Fr5); in milk instead of water was added to the basic formula (Gl-Fr6 ) dough was prepared

from a premix market(corn starch, starch wheat gluten free, sugar, guar gum, sweeteners

grapes, wheat gluten free cellulose, pectin, emulsifiers - mono and diglycerides of fatty acids,

Hungary)(Gl-Fr7); 35% potato pulp Laura, red potato variety (Potato Research and

Development Station Târgu Secuiesc, Covasna, România) at the mater formula addiding(Gl-

Fr8). Mixing was done with the mixer type SADKIEWICZ (Poland). Add all ingredients

together. Xanthan was hydrated by mixing for 20 minutes before, by 50 % of the total amount

of recipe water. The kneading temperature was 28 °C, during 10 minutes. The fermentation is

performed at 30 °C for 50 minutes and baking for 30 minutes at 240 °C. Shall be used for

baking trays with size (17 x 9.5 x 7.5) cm.



72

Preparing the potatoes pasta: PP (potato pasta) was obtained by hydro thermal processing of

the unpeeled raw potato for 30 min. at water boiling temperature, then cooling it, peeling, and

mashing it by passing it through the 2 mm mesh sieve (Iancu ML., et al., 2010).

The baking test were carried aut in an electric oven with an incorporated proofing chamber

(type ESM3710 SADKIEWICZ- Poland). Bread was analyzed after 2 hours from the removal

from the oven. Determination of specific volume in cm3/g was made by displaced volume

method of rape seed (AACC-2001), the porosity is in principle to determine the total volume

of voids in a given volume of the crumb, knowing the volume of oil displaced crumb

scoreless and its weight (SR 91-2007), elasticity, humidity and baking losses on.

Result and discussion

Physico-chemical analysis results of gluten-free bread are shown in Table 1.

Table 1. Values primary, quality indicators of bread gluten free

Gl-Fr1 Gl-Fr2 Gl-Fr3 Gl-Fr4 Gl-Fr5 Gl-Fr6 Gl-Fr7 Gl-Fr8

Vs[cm3·1g-1] 2,01 2,46 1,64 1,67 2,34 1,94 1,98 2,39 aP [%] 73,4 69 66 73,4 75,2 86 83 75 bE [%] 88 90 80 88 91,2 89 89,7 90 cu [%] 41,6 50,2 51,2 55,1 43 40,2 43,5 45 dPc[%] 13,9 17,4 6,67 6,7 10,2 13 14 16,5

a-porosity; b-elasticity; c-moisture crumb; d- baking losses

The control sample of bread (control sample) (Gl-Fr1) has a remanence of odor (yeast). The

kernels have a hard texture and is brittle. Shell color is pale white dusty appearance. To

overcome these drawbacks has changed the recipe. Thus by the egg, core is soft and crumbly

baked egg taste is pleasant, popcorn. The crust is tender white uniform. The sample was loose

biochemically did not smell of yeast (GI-Fr3).

The egg product contributed to a more airy. Improved nutritional value, basic recipe is a

minimalist recipe was found that on it can create a new recipe to remove deficiencies. Bread

texture is an important quality parameter. Supplementation of gluten free bread with various

types of plant flour was done in other studies, and in this way affect crumb texturre (Korus

J., et al., 2015).



73

In this study, the bread has to chewing texture is rough, with remanence in the mouth,

crumbly is great. This drawback has been removed through the addition of substances

improving (mono and diglycerides of another flour, milk, egg). They are expensive materials,

which in turn give intolerance sometimes and some are not very natural (Gl-Fr 6,7). The milk

is used for making bread without gluten. It has high biological value and contains selenium

(8μg) where cereals are deficient (Sager M., 2006).

Figure 1. Macroscopic characteristics for gluteh free breads prepared from different materials: 1- Gl-Fr1;

2- Gl-Fr1 and Gl-Fr3; 3- Gl-Fr4 and Gl-Fr5 ;4- Gl-Fr6 and Gl-Fr7 ; 5- Gl-Fr8

Was attempted addition of 35% PP (red potato variety Laura). There were obtained results of

the macroscopic sample, GI-Fr8, similar to those of the reference sample the GI-Fr1 (Figure

1). We managed to remove crumb texture roughness and crumbly. Values determine quality

indicators are presented in Table 1. The specific volume of the samples had the highest value

for the sample with egg and yeast (Gl-Fr2) and by 35% by replacing of the starch with PP



74

(Gl-FR8). Porosity and elasticity for these samples have been improved. Baking losses are,

however, higher (GI-Fr8) as compared to control sample (GI-Fr1), from 13.9% to 16.5%.

Chemical loosening given good results determine quality indicators, but there is no specific

flavor of bread.

Conclusions

Partially replaced with PP (35% - Laura - GI-Fr8) corn starch and thus decrease the

concentration of xanthan gum had great influence on the firmness of the core structure. It was

shown that supplementation with PP and other material has a positive impact on the specific

volume of the products of elasticity and moisture core. As prospects for further research will

extend this short communication studies by choosing a variant, the cheapest.

References

AACC International Method 10-05(2001): Guidelines for Measurement of Volume by Rapeseed Displacement Alvarez-Jubete L., Arendt EK., Gallagher E., Nutritive value of pseudocereals and their increasing use as

functional gluten-free ingredients. Trends Food Sci Technol, 2010, 21, 106–13.

El-Chammas K., Danner E., Gluten-free diet in nonceliac disease. Nutr Clin Pract, 2011, 26:294–299.

Iancu Maria Lidia, Iancu Bianca Maria, Quality assurance in the gluen-free bread technology, using responce

surfaces method, Journal of Agroalimentary Processes and Technologies, 2015, 21(1), 14-20,

http://journal-of-agroalimentary.ro

Iancu M.L., Luiza M., Haubelt, G., Experimental model for the application of the flourgrafic technique to the

study of the mixture of flour and raw potato, or hydro thermally processed potato, Journal of

Agroalimentary Processes and Technologies, 2010, 16(1), 41-48.

Korus Jarosław, Mariusz Witczak, Rafał Ziobro, Lesław Juszczak, The influence of acorn flour on rheological

properties of gluten‑free dough and physical characteristics of the bread, Eur Food Res Technol, 2015,

doi:10.1007/s00217-015-2417-y.

Matos M.E., Rosell CM., Understanding gluten-free dough for reaching breads with physical quality and

nutritional balance. J Sci Food Agric., 2015, 15, 653-661, doi:10.1002/jsfa.6732.

Matos Segura ME., Rosell CM., Chemical composition and starch digestibility of different gluten-free breads.

Plant Foods Hum Nutr, 2011, 66(3):224–230.

O’Shea N., Arendt E., Gallagher E., State of the art in gluten- free research. J Food Sci, 2014, 79(6): R 1067–

R1076.

Padalino L., Mastromatteo M., Lecce L., Cozzolino F., Del Nobile MA., Manufacture and characterization of

gluten-free spaghetti enriched with vegetable flour. J Cereal Sci, 2013, (57) 333-342. doi:

10.1016/j.jcs.2012.12.010



75

Sager M., Selenium in agriculture, food and nutrition. Pure Appl Chem , 2006,78:111–133

Sharma M.G., Pereira M., WilliamsK. M., Gluten detection in foods available in the United States – A market

Survey, Food Chemistry 169 (2015) 120–126.

SR 91(2007): Pâine și produse proaspete de patiserie.



76

DECONTAMINATION KINETICS OF AFLATOXIN M AT DIFFERENT

DOSES OF UV RADIATION Ketney Otto*1

1“Lucian Blaga” University of Sibiu, Faculty of Agricultural Sciences, Food Industry and Environmental

Protection, Sibiu, Romania , (* Corresponding author: e-mail: [email protected])

ABSTRACT:

Aflatoxin M1 (AFM1) is a metabolite that appears in milk and dairy products from animals fed with aflatoxin

B1 (AFB1) contaminated plants. These aflatoxins are highly toxic and have a high degree of carcinogenicity.

Given that the AFM1 molecule is thermostable and cannot be destroyed by pasteurization or UHT, this

research aims to study kinetics of UV-induced photodegradation of AFM1 from artificially contaminated milk

samples at a concentrations twice the allowed level by Regulation (EC) no. 1881/2006. The kinetics of the

reaction indicates that a radiation dose of 500 μW cm-2 is effective within 5 seconds of exposure. Knowledge

of these data is important because this process shows appropriateness for scaling-up to industrial level.

Keywords: aflatoxin , milk, photodegradation , irradiation

1. Introduction

Mycotoxins are toxic compounds produced by fungi as secondary metabolites. The

most important groups of mycotoxins in foods are aflatoxins, ochratoxins, trichothecens,

zearalenone and fumonisins (Bennett and Klich, 2003). The molds have the potential to

produce mycotoxins and the process is greatly influenced by environmental factors like

temperature, relative humidity, insect damage, drought and poor storage conditions (Miraglia

et al., 2009; Prandini et al., 2009).

Aflatoxin is most prevalent in tropical and subtropical regions cultures of the world

(Strosnider et al., 2006) . Aflatoxins are the most studied mycotoxins, mainly consisting of

four natural compounds: aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1) and

aflatoxin G2 (AFG2) (Fallah, 2010). Aflatoxin M1 (AFM1) and M2 (AFM2) are hydroxylated

metabolites of aflatoxin B1 (AFB1) a B2 (AFB2) and can be found in dairy products from

animals that have ingested contaminated feed (Unusan, 2006). Although AFM1 is the main

monohydroxylated derivative of AFB1, it is less carcinogenic and causing genetic mutation



77

than AFB1, however has a high level of genotoxicity and represents a health risk (Heshmati

and Milani, 2010).

Milk and dairy products are essential for human consumption. AFM1 levels in milk

shows increasing attention because of their human health repercussions. Most of AFB1

consumed by feed is degraded in the rumen of dairy cows, but an important part is resorbed

and metabolized rapidly in the liver AFM1 by cytochrome P450. AFM1 is relatively stable

and circulates in the blood until it is secreted in milk and bile or excreted in the urine

(Prandini et al., 2009). AFM1 is relatively stable during pasteurization, sterilization,

preparation and storage of dairy products (Wiseman and Marth, 1983)

To protect the health of humans and animals, over 100 nations have established

maximum tolerable levels of aflatoxin in food (Wu and Guclu, 2012). Currently, the presence

of aflatoxin in feed, milk and dairy products can be efficiently monitored in Europe and other

developed countries, so the European Community has accepted for AFM1 of 0.050 μg Kg−1 in

milk (European Commission, 2001)

Because the problem of aflatoxin contamination is a potential hazard, developing

effective methods of detoxification aflatoxin degradation becomes an important approach

(Akbas and Ozdemir, 2006)

Some of the mycotoxin decontamination methods literature include: physical (Fan et

al., 2013; Liu et al., 2011; Liu et al., 2010; Park et al., 2007) , chemical (Adamovic et al.,

2014; Carraro et al., 2014; He et al., 2014; Ismaiel and Tharwat, 2014; Luo et al., 2014; Luo

et al., 2013) or biological (Alberts et al., 2009; Caloni et al., 2006; Cheng et al., 2010;

Corassin et al., 2013; Elsanhoty et al., 2014; Gratz et al., 2004; Haskard et al., 2001; Hathout

and Aly, 2014; Samuel et al., 2014; Schatzmayr et al., 2006; Vijayanandraj et al., 2014)

methods. However, the methods are not generally applicable, efficient and practical (Peltonen

et al., 2000).

Aflatoxin B1 absorbs ultraviolet (UV) at 222, 265 and 362 nm, and the highest

absorption is 362 nm. Irradiation at 362 nm activates the susceptibility to degradation.

However, the degradation process is effective at a pH less than 3 or greater than 10 (Lillard &

Lantin, 1970). Li and Bradley Jr (1969) demonstrate that the rate of degradation depends on

the film thickness and the depth of diffusion of the rays. Since aflatoxin susceptibility to UV

radiation is increased we proposed in this paper to study the kinetics of degradation of

aflatoxin M1 from milk at different doses of ultraviolet (UV) of various energies. This paper

is a continuation of the previously performed by (Ketney et al., 2010).



78

2. Materials and methods

2.1. Sample preparation

Samples of raw cow milk come from many milk manufacturers. The milk was

purchased from the market and it was tested for presence of residual aflatoxin M1 before the

start of the experiment. Milk temperature during the experiment was 23 0 C

2.2. Samples contamination

A 10 µg mL-1 solution of AFM1 (Sigma – St. Louis, MO) in a methanol/chloroform

mixture was prepared and kept frozen at –200C. Because AFM1 is photosensitive the

solutions were stored in aluminum foil-wrapped vials, and were not exposed to daylight or

placed close to fluorescent light sources. The solution of AFM1 was added (under continuous

stirring) to each sample to provide a calculated concentration of AFM1 in the milk of 0.1 ppb

(100 ng L-1) which is twice the maximum permitted level by EC (2006). The samples were

store frozen in the dark until used.

2.3. Photodegradation procedure

The irradiation procedure, device irradiation and irradiation conditions has been

described elsewhere (Ketney et al., 2010). In this paper we used 50 mL of cow's milk were

introduced in the irradiation device between two plates having 5 mm distance between them

(no air bubbles) and was subjected to UV radiation at a wavelength of 253.7 nm for 45

seconds

2.4. Analysis o aflatoxin

Extraction of AFM1 and ELISA test procedure were performed according to the instructions

of the test kit (Ridascreen Art No.: R1101, R-Biopharm).

2.5. Statistical analysis

All analytical measurements were done in triplicate. Values of different parameters were

expressed as the mean ± standard deviation. Statistical examination was down using the IBM



79

SPSS Statistics version 21 packet program for Windows (SPSS, Chicago, IL, USA). The

study of variance (ANOVA) was performed on the obtained data.

2.6. Mathematical model Description

The inactivation process is described by a relatively simple mathematical description of

the first order inactivation kinetics of AFM1. UV radiation depends on the rate constant k and

the mathematical model involves changes in the rate of inactivation of aflatoxin by UV

radiation dose.

For this study we followed the assumed first order reaction kinetics for the process of

aflatoxin inactivation and the experimental data were obtained for a 5 mm milk width

between the plates.

Knowing the concentration in each test and the molecular mass, the number of moles of

aflatoxin was calculated.

3. Results and discussion

AFM1 content in milk samples was below the detection limit (0.005 μg L-1), and after

artificial contamination was AFM1 content (100 μg L-1). AFM1 content after irradiation is

shown in Table 1.

Table 1 Changes in the concentration of AFM1 in various doses of irradiation (5 mm milk width between plates)

Time s

DOSE* 1 UV ± SD** ng L-1

DOSE* 2UV ± SD**

ng L-1



0 99.8 ± 0.2 99.9 ± 1.4 99.7 ± 0.5 99.8 ± 0.2 5 94,2 ± 2.8 89,6 ± 3.2 81,7 ± 2.2 74,6 ± 0.1 10 88,2 ± 0.2 81,3 ± 2.2 72,3 ± 0.2 63,2 ± 0.7 15 83,3 ± 0.1 74,5 ± 0.1 63,1 ± 2.1 54,3 ± 0.5 20 78,3 ± 2.2 68,5 ± 3.2 57,2 ± 2.3 47,2 ± 1.2 25 73,3 ± 0.2 61,4 ± 0.7 51,1 ± 1.1 40,8 ± 2.3 30 68,4 ± 2.3 57,5 ± 1.2 46,2 ± 0.4 36,3 ± 1.6 35 64,1 ± 0.5 53,6 ± 0.2 43,1 ± 1.7 33,2 ± 0.4 40 61,3 ± 0.5 49,4 ± 0.5 39,2 ± 0.5 30,8 ± 2.5 45 58,6 ± 1.2 45,7 ± 0.6 37,8 ± 1.8 28,5 ± 1.9

DOSE* - DOSE 1 = 125 μW cm-2, DOSE 2 = 250 μW cm-2, DOSE 3 = 375 μW cm-2 and respective DOSE 4 = 500 μW cm-2 SD** - Standard deviation for three determinations



80

The lowest value of AFM1 was 28.5 ng L-1, at the radiation DOSE 500 μW cm-2, and the

highest was 99.9 ng L-1 at the radiation DOSE 250 μW cm-2. There is significant difference

from statistical point of view between radiation doses (p <0.05) and the highest variable

content of AFM1 was inactivated at the DOSE of 500 μW cm-2 irradiation. A highly

significant difference (p <0.001) was obtained for an irradiation time of 25 seconds.

Variation of AFM1 at different irradiation doses is observed at 45 s irradiation time: at a

DOSE of 500 μW cm-2 the extent of inactivation was much stronger compared to the

inactivation obtained by a DOSE of 125 μW cm-2.

The model described above implies that the rate of degradation has been associated with

AFM1 concentration. The calculation order of the reaction rate constant k was done by using

the MathCAD program 14 in a similar manner to that described by (Silivia et al., 2002).

The first order kinetics assumption for the aflatoxin inactivation was checked by graphical

representation of ln (CA) = f (t) (Figure 1).

In each case an approximate straight line dependence was obtained, by thus verifying the

reaction order by the value of the slope, and therefore the reaction order is one. The rate

constant k was obtained from the slope of the straight line regression for each case.

This study provided evidence of effectiveness on reducing milk AFM1 at a level of

0.050 μg Kg-1 accepted in accordance with European Union (Carraro et al., 2014)

Figure 1. Graphical representations of the changes in the concentration of AFM1 in various

doses of irradiation for UV dose, (DOSE 1 = 125 μW cm-2, DOSE 2 = 250 μW cm-2, DOSE 3 = 375 μW cm-2 and respective DOSE 4 = 500 μW cm-2).



81

There are different studies (Liu et al., 2010; Yousef and Marth, 1986; Zhu et al.,

2013) in regard to the UV radiation process of the mycotoxins and photodegradation kinetics

in different environments However, degradation products AFM1 of UV photons are still

unknown. However, based on previous paper (Liu et al., 2010) of AFB1 on photodegradation,

structural formulas have been proposed for these products of degradation. These formulations

are based on exact mass calculated from the molecular formula and MS-MS fragmentation

formation provided by Q-TOF.

A study similar to the photodegradation kinetics of AFB1 performed Aibara and

Yamagishi (1968) demonstrated for the first time first-order kinetics, as a result of one of the

intermediate reaction products of photodegradation may be a coumarin molecule dimerized.

However Falguera et al. (2011) demonstrated that intermediate products products have same

toxicity, and similar research this should be extended in this regard.

4. Conclusions

Overall cost of the problems caused by aflatoxin is high - both in terms of losses

related to the sale of products contaminated with aflatoxin and human health problems

because of the negative effects of exposure to aflatoxin.

UV irradiation method was applied to study the kinetics of photodegradation AFM1.

Results of this study provide detailed information on the changes in the kinetic stability of

AFM1 in milk for a short time (45 s). The mathematical model experimentally obtained in

this study provides valuable information on kinetic parameters of the process of milk

treatment with UV in order to inactivate the AFM1 molecule AFM1 in optimum UV

irradiation condition. The data show that degradation of AFM1 in milk corresponds to a first

order reaction. Changes in the rate of degradation in relation to the dose of irradiation suggest

that an irradiation DOSE of 500 μW cm-2 is more efficient. However further studies are

needed on the identification of degradation compounds and their toxicological and long-term

effects on medium.

In conclusion, the current study demonstrated that UV radiation at 253.7 nm could be

an effective natural method of reducing the levels of AFM1 in milk. AFM1 degradation

kinetics was consistent with a first-order reaction model.



82

5. Acknowledgements

This work was supported by the strategic grant POSDRU/159/1.5/S/133255, Project ID

133255 (2014), co-financed by the European Social Fund within the Sectorial Operational

Program Human Resources Development 2007 – 2013.

6. References

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Aibara, K. and Yamagishi, S., Effects of ultraviolet radiation on the destruction of aflatoxin B1. In: U.j.p.o.t.m.a.U.S.D.o. Interior (Ed.), Proceedings of the first US-Japan conference on toxic microorganisms, Washington DC, Effects of ultraviolet radiation on the destruction of aflatoxin B1.

Akbas, M.Y. and Ozdemir, M., Effect of different ozone treatments on aflatoxin degradation and physicochemical properties of pistachios. Journal of the Science of Food and Agriculture, 2006, 86: 2099-2104.

Alberts, J.F., Gelderblom, W.C., Botha, A. and van Zyl, W.H., Degradation of aflatoxin B(1) by fungal laccase enzymes. Int J Food Microbiol, 2009, 135: 47-52.

Bennett, J.W. and Klich, M., Mycotoxins. Clinical Microbiology Reviews, 2003, 16: 497-+. Caloni, F., Stammati, A., Frigge, G. and De Angelis, I., Aflatoxin M1 absorption and cytotoxicity on human

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V. and Tateo, F., Clay minerals as adsorbents of aflatoxin M1 from contaminated milk and effects on milk quality. Applied Clay Science, 2014, 88-89: 92-99.

Cheng, B., Wan, C., Yang, S., Xu, H., Wei, H.U.A., Liu, J., Tian, W. and Zeng, M., Detoxification of Deoxynivalenol by Bacillus Strains. Journal of Food Safety, 2010, 30: 599-614.

Corassin, C.H., Bovo, F., Rosim, R.E. and Oliveira, C.A.F., Efficiency of Saccharomyces cerevisiae and lactic acid bacteria strains to bind aflatoxin M1 in UHT skim milk. Food Control, 2013, 31: 80-83.

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84

PHYSICAL AND GEOGRAPHICAL PARTICULARITIES OF

VINEYARDS IN THE APOLD DEPRESSION

Ecaterina Lengyel

”Lucian Blaga” University of Sibiu, Romania , Bl. Victoriei no 10 e-mail [email protected]

ABSTRACT The relief in the Apold Depression is the most dynamic, through its accentuated fragmentation and slope

morphodynamics, aspects that appeared from instability and the presence of current associated

geomorphological processes. Climatic elements are reflected in the daily and seasonal dynamics of the

general atmospheric circulation. The hydrography dynamics is less visible, as it is generally noticed based on

slow shifts of stream beds taking place in main valleys, including in those introduced by humans. All these

elements contribute to customizing the viticultural geographic area Apold.

Keywords: Apold Depression , lithology, water resources

The soils and edaphic coating of the Apold Depression are mosaical, resulting from a

combination of several factors: lithology (alternation of marlstone, sand, sandy clay, gravels

and gypsum), its fragmentation and stage layout, with the contribution of the depression

climate which has oceanic influences, the massive deforestations (so as to use the land for

agriculture), which led to the modification of soils: from sylvester soils to Secaș leached

chernozems and pseudorendzinic soils (Teușan-Pleșia 2011). Gradual and cvasi-horizontal

surfaces led to the conservation of soils (Buza 2005, ). They are presented as evolved soils

and are categorized as follows:

- in river wet meadows: gleysols and alluvial soils which will be exploited as arable land

for numerous crops, especially for industrial plants and vegetables (as they are fertile), as

well as for grassland;

- on terrace treads and glacis: Secaş leached chernozems (typical cambic and moderate

leached chernozems and typical phreatic-wet cambic chernozems) which are rich in

humus and good for cultivating cereals;



85

- foothills are characterized by holoacid luvic brown and superficially oligo-alkaline

sylvester soils;

- on the interfluves between the Câlnicului and Reciului, Dobârcăi and Gârbovei valleys,

we find pseudogleised luvic brown soils (partially covered by Carpinus betulus and

Quercus petraea) which are less fertile, thus favoring natural pastures. On sunnier slopes,

they support tree and fruit and wine-growing lands. Pseudorendzinic and rendzina soils

(which have a well-developed humus layer, are used for pastures and grassland and, in

some places, even bare rocks can be found on slopes affected by landslides and ravines.

As a consequence, we can determine the physical and geographical particularities of the

geographical area in the Apold Depression based on the dynamics of its constituent parts

(Ciobanu 2002, Buza et al. 1985, Badea et al. 1975).

Water resources in the Apold Depression

The Apold Depression is part of the Mureș River basin (Lower Corridor of the Mureș River).

Through the way it flows and its position, together with the lithology and climatic conditions,

it ensures a permanent regime of the river system. The Transylvanian pericarpathian supply

comes from rain and snow (winter flow is higher than summer flow; the highest flow is

recorded in March, even though April is richer in more frequent precipitations). The annual

flow is directly influenced by the thickness of the snow layer (the duration and the intensity

of its melting, its stability) and by the relief.

Surface waters: In the Apold Depression, the water system is made up of two collecting

arteries: Secașul Mare and its tributaries together with the Petrești reservoir on the Sebeș

River and the Sebeș River and its tributaries.

The Sebeș River represents the West border of the Apold Depression; it is a left tributary to

the Mureș River, springing from the Southern slope of the Cindrel Mountain, close to the

Șteflești col, at an altitude of 2100 m, being 96 km long and having a basin surface of 1289

km2; between the localities Lancrăm and Sebeș, it forms a very big alluvial fan due to the

erosion and transport force. Secașul Mare is the most important tributary in the lower corridor

and lies at North-West of Lancrăm, springing from an altitude of 450 m and draining at 235

m. The basin has a surface of 567 km2, is 42 km long and the multiannual average flow rate is

rather low (under 1 m3/s). Most of the left tributaries (Dobârca, Apold, Gârbova, Pustia,

Răhău, Câlnic) flow into Secașul Mare River, which is why the river basin is visibly

asymmetric and more developed on the left side. In 2005, Costea Mărioara identified two

hydrographic convergence sectors in this area and called them "markets where waters



86

gather": the Cunţa - Cut sector towards which Boz and Şpring, Valea Satului, Câlnic, Răhău

and Valea Caselor converge and the Apoldu de Jos - Miercurea Sibiului sector towards which

Apold, Dobârca, Pustia and Gârbova converge.

Figure 1. Sebeș river

They originate in two rivers, i.e. Secașelor flowing from Luduș (Secașelor Plateau) and

Apold collecting most its water from the right slope of Furcilor Hill (rivers flowing from and

the Cindrel Mountains); the rivers on the right contribute to the flow rate insignificantly.

The hydrographic map of the Apold Depression present:

- swamp sectors generally situated close to the main confluences (Aciliu with Amnaş,

Apold with Secaşul Mare, Gârbova with Reciu, Boz with Secaşul Mare etc.);

- confluence sectors (some of them as a series: Apold, Dobârca, Pustia, Gârbova,

Câlnic, Răhău, Valea Caselor) and of torrential convergence;

- unplaiting sectors (on the lower corridor of the Sebeș Valley);

- mineral springs of various chemical compositions (Miercurea Băi, Cunța, Cut);

- river planning for hydropower (Petrești) or for pisciculture (Apold, Boz and

Slatina/Daia – in the case of the last two, in the middle sectors, outside the depression,

Bâscă, 2009)



87

Figure 2. Petrești lake

- meandering sectors on the Secașul Mare;

Figure 3. Secașul Mare river

There are anthropological stream bed shifts on the Sebeș river. The studied area presents two

hydrometric stations, i.e. Secașul Mare-Cunța and Sebeș-Petrești, which is marked by human

activity through a dam reservoir.

Groundwaters

Due to sedimentary rocks (clays, malrstone, sands, gravels), ground waters are an aquiferous

complex of piedmont deposits with aquiferous layers situated at depths between 10 and 15 m,

have an alkaline pH of 7.4 and are a pale shade of yellow.



88

Figure 4.Sedimentary rocks comprising gravels, clay and sands

As the clay-sand or sand formations include gravels, drinking water springs have a high

density and some springs on the Câlnic - Dobârca course are slightly mineralized.

The groundwater reservoirs are less represented in the peripheral monocline of the Secașelor

Plateau. They are more developed in the miocene deposits as an aquiferous complex of clay

and marlstone which is highly mineralized. Following shallow water drilling and drilling in

the waters collected in the basins of the Miercurea Sibiului baths, the chemical composition

of Sodium chloride ranged between 19.3 and 40.2 g/l Cl–Na48; they are subsequently

enriched with I-, Br-, Ca++ and NH4+, to which sodic chlorinated water from Cunța and Cut

are added, enriched with other chemical elements (Bogdan, 2002)

Figure 5. The sodic chlorinated waters collected in the basins of the Miercurea Sibiului baths



89

The wines in the Sebeş-Apold Vineyard, Traminer Roz, Pinot Gris, Muscat Ottonel,

Sauvignon, Chardonnay, Neuburger, Rhine Riesling, Italian Riesling, Fetească Regală,

Fetească Albă bear the imprint of type, soil, microclimate, poultry farmer and winemaker and

are characterized by: fruitiness, freshness and a slightly higher acidity. They are fine wines,

with a wide range of aromas that are typical and specific to their variety.

ACKNOWLEDGMENT This work was supported by the strategic grant POSDRU/159/1.5/S/133255, Project ID 133255 (2014), co-financed by the European Social Fund within the Sectorial Operational Program Human Resources Development 2007-2013

REFERENCES Bâscă I. (2009), Petreşti –şapte milenii de istorie. Repere monografice, Edit. Altip, Alba Iulia.

Bogdan Octavia (2002), Potenţialul termic al judeţului Sibiu, premise pentru dezvoltarea activităţilor turistice,

în Rev. Geo-Carpathica, II, 2, Edit. Universităţii „Lucian Blaga”, Sibiu.

Badea L., Sandu Maria (1975), Profil geomorfologic prin depresiunile Apoldului şi Săliştei, SCGGG, Geogr.,

XXII, Edit. Academiei, Bucureşti.

Buza M. (2005), Noul sistem român de taxonomie a solurilor, în Rev. Geografică, T. XI, 2005, Serie Nouă,

Academia Română, Institutul de Geografie, Bucureşti.

Buza M. (2010), Oiconimele din depresiunile Sibiului şi Făgăraşului – reflectare fidelă a condiţiilor geografice

şi istorice, în Rev. Geo-Carpathica, X, 10, Edit. Univ. „Lucian Blaga”, Sibiu.

Buza M., Hozoc I. (1985), Valea Sebeşului, Edit. Sport-Turism, Bucureşti.

Ciobanu Rodica (2002), Depresiunea Sibiului. Studiu geologic, Edit. Univ. „Lucian Blaga”, Sibiu. Dinamica

spaţiului geografic în Depresiunea Apoldului

Teușan-Pleșia, (2011), Dinamica spațiului geographic în Depresiunea Apoldului, Edit. Univ. „Lucian Blaga”, Sibiu.



90

EVALUATION OF ASCORBIC ACID AND TOTAL ANTHOCYANINS

IN FRESH CHEESE ENRICHED WITH BILBERRY, CRANBERRY

AND DRIED BERRIES

Mihaela Amăriuţei, Mihaela-Adriana Tiţa, Simona Oancea

Department of Agricultural Sciences, Food Industry and Environmental Protection, „Lucian Blaga” University

of Sibiu, Sibiu , Romania

Abstract The aim of the present paper was to evaluate the content of antioxidant compounds, such as ascorbic acid

and total anthocyanins in fresh cheese enriched with fruits. Three types of fresh cheese enriched with fruits

of bilberry, cranberry and dried berries mixture as tea, were investigated in comparison to control

(untreated). The sensory characteristics, the content of ascorbic acid and total anthocyanins was monitored

during 14 days of storage at 2-4oC. Quantitative determination of ascorbic acid showed highest values for

fresh cheese with added cranberries. The content shwed similar values during storage for all samples.

Regarding the content of total anthocyanins, significant results were found in case of fresh cheese with

added bilberries.

This study showed that the shelf life of fresh cheese could be extended by at least one day, given the good

sensory characteristics after 15 days of refrigerated storage. Beneficial antioxidant effects determined by the

presence of ascorbic acid and anthocyanins from added fruits was registered in the investigated cheese.

Keywords: fresh cheese, berries, ascorbic acid, anthocyanins

INTRODUCTION

Varied textures of fermented and non-fermented cheese is mainly composed of the casein

matrix containing fat, variable amounts of lactose, minerals and vitamins. (Banu C., 2009)

Curd is one of the most prevalent types of fresh cheese consumed in Romania. Fresh

cheese is produced by processing the skimmed or whole cow's milk, leading to a dairy

product with high nutritional value and dietetics. Fresh cheese is a good source of fat,

calcium, folic acid, vitamins A and D, giving approximatevly 25% of the protein daily intake.

However, it lacks some vitamins, such as ascorbic acid (vitamin C). Such vitamins are

responsible for reducing free radicals and consequently improving degenerative diseases.

(Jacob R.A. et al, 2002) Addition of vitamins proved to have a beneficial effect both on the

human body and on the compositional integrity of the food product.



91

Ascorbic acid is responsible of producing neurotransmitters, playing a key role in

carbohydrates metabolism, formation of colagen, absorption of ferrum, folic acid and some

amino acids, and scavenging of free radicals. (Combs G.F., 2001)

Addition of some other antioxidant compounds, such as other vitamins or anthocyanins, in

food products may determine health benefits (Țifrea A. Et al, 2012; Klein A.D., 2014).

Anthocyanins are water soluble pigments responsible for red, blue colors of fruits, mainly.

They display wide positive health effects in particular regarding chronic diseases, based on

their strong antioxidant activities. These bioactive compounds found large application in

pharmaceutical and food industries. Their molecular stability under different conditions

(temperature, pH, presence of enzymes, etc.) represents a challenge for such application in

particular in food products. (Oancea S. et al, 2013; Oancea S. et al, 2015)

It is well known that health depends on biological and environmental conditions, and also

on culinary habits of consumers. Therefore it becomes very important to completely

characterise plant antioxidant compounds, in order to use them as ingredients to produce new

food products (nutraceuticals).

The chemical structures of the most important antioxidant compounds (ascorbic acid, anthocyanins) are presented in Figures 1-2.

Figure 1. Chemical structure of ascorbic acid

Figure 2. General chemical structure of anthocyanidines.



92

The aim of the present paper was to improve the nutritional value of fresh cheese by

adding three type of ingredients rich in ascorbic acid and anthocyanins (frozen bilberries,

dried cranberries and dried berry mixture tea). Sensory characteristics, pH and acidity, the

content of ascorbic acid and total anthocyanins have been evaluated during 14 days of storage

at 2-4 ° C.


Biological samples and reagents: fresh cheese was obtained from fresh whole cow's milk

which was subjected to the following: sensory analysis, physico-chemical analysis,

pasteurization at 85ºC for 20 minutes, cooling setpoints until cooling reached 35ºC,

renneting, curd processing (Tița M.A., 2002). Curd (fresh cheese) was divided into four

samples, separately treated with: (1) control samples (without addition); (2) dried cranberries

(25 g fruits 100g-1 FM cheese); (3) dried berries mixture tea (2 g fruits powder 100g-1 FM

cheese), and (4) frozen bilberries (25 g fruits 100g-1 FM cheese). The samples were stored in

tapped containers for 14 days at 2-4oC. Analysis of ascorbic acid and total anthocyanins

content was carried out in the 1st, 7th and 14th days of storage.

Chemical reagents of analytical purity were used in all experiments.

Physico-chemical analysis: Physico-chemical analysis of milk as raw material was done

using EKOMILK TOTAL instrument. Acidity was determined by titration method, while pH

was measured using the pHmeter.

Assay of ascorbic acid: the content of ascorbic acid was determined using the iodometric

titration procedure (Izuagie A.A. at al, 2007). Measurements were done in three replicates.

The content of ascorbic acid was expressed as mg 100g-1 FM.

Assay of total anthocyanins: anthocyanins extracted from samples with 96 % ethanol for 1

hr., was spectrophotometric ally determined by the pH differential method. (Wrolstad R.E.,

2001) The CECIL CE1021 1000 Series spectrophotometer was used. Measurements were

done in three replicates. Total anthocyanins were expressed in mg 100g-1 FM.



93


The analysis of milk as raw material, in terms of sensory and physico-chemical parameters,

showed the quality of milk used for obtaining fresh cheese, as presented in Table 1.

Table 1. Quality of milk used for preparation of fresh cheese, as determined using Ekomilk

Total instrument. Parameter Value

FAT, % 4.72

SNF, % 8.5

DEN 27.25

PROT, % 3.22

FP 57.25

T, ºC 20.5

LAC, % 4.65

Z, Ms / cm 2.2

pH, R 6.1

Three types of fresh cheese enriched with fruits high in antioxidants were obtained, as

follow: 1 – control, 2 – fresh cheese with dried cranberries, 3 – fresh cheese with dried

berries mixture tea, and 4 – fresh cheese with frozen bilberries.

The results regarding the content of ascorbic acid in the studied fresh cheese samples are

shown in Figure 3. As observed, the highest content was found in sample 3, as cranberries are

fruits rich in ascorbic acid.

The content of total anthocyanins in the studied samples highly varied among samples.

Sample 4 with bilberries showed relevant amounts in the final dairy product. Compared to

cranberries addition, bilberries one is recommended as these fruits were added in the

fresh/frozen form, while cranberries were added in dried form which did not permit an

optimum migration of such compounds in the product. However, as shown in Figure 4, the

content slightly decreased during the 14-days period of storage at 2-4oC.



94

Figure 3. The content of ascorbic acid in the three enriched fresh cheese samples during storage.

Figure 4. The content of total anthocyanins in fresh cheese enriched with bilberry fruits (sample 4) during

storage. Other authors reported different retention of antioxidant compounds, such as catechin,

epigallocatechin gallate, tannic acid, green tea extract, added to cheese curds or green tea

extract added to Cheddar-type cheese in order to obtain better quality and functionality dairy

products with improved health benefits (Han J. et al, 2011; Giroux H.J. et al., 2013).

The change in lactic acid formed during fermentation studied over the 14-days storage

period is presented in Table 2.

The pH values were similar during the storage period for each sample, except of control in

which pH increased, as shown in Table 3.

Table 2. The change in lactic acid in studied fresh cheese samples.

Sample Lactic acid during storage (days) 0 1 7 14

1 0.702 0.747 0.792 0.855

2 - 0.720 0.774 0.810



95

3 - 0.765 0.810 0.819

4 - 0.720 0.765 0.792

The change of total acidity in the investigated fresh cheese samples is presented in Figure

5. Sample 1 showed an initial acidity of 78ºT, and a final value of 95ºT, being the highest

value among all samples. The increase in total acidity might be due to the physico-chemical

degradation of products or microbial actions. Sample 4 showed minimum change in total

acidity during the 14-days period of storage. Table 3. Change of pH during storage period in studied fresh cheese samples.

Sample pH during storage (days) 0 1 7 14

1 4.0-4.5 4.5-5.0 4.5-5.0 5.0-5.5 2 - 4.0-4.5 4.0-4.5 4.0-4.5 3 - 4.5-5.0 4.5-5.0 4.5-5.0

4 - 4.5-5.0 4.5-5.0 4.5-5.0

Figure 5. Change of total acidity in studied fresh cheese samples during storage.

CONCLUSIONS

Three types of fresh cheese enriched in fruits high in antioxidant compounds were studied in

terms of quality and content of bioactives. The highest content of ascorbic acid was found in

fresh cheese enriched with dried cranberries. The highest content of total anthocyanins was

found in fresh cheese with bilberry. During the 14-days storage period, total anthocyanins

slighlty degraded.

Regarding the pH, an increase of the value was found in control sample, in which physico-

chemical and sensory degradation started, compared to the other samples, preserved through



96

addition of fruits rich in antioxidant compounds. The cheese enriched in such compounds

maintained quality characteristics during longer periods.

Regarding the total acidity, it was maintained at similar values in fresh cheese enriched

with bilberry showing better stability during storage period. Cheese acidity may cause both

physical and chemical quality degradation.

The obtained results showed that additon of natural antioxidant compounds from berries

may lead to longer shelf life periods for fresh cheese.

REFERENCES Banu C., Tratat de industrie alimentară, Tehnologii alimentare, Editura ASAB, Bucuresti, 2009.

Brouillard R., Analysis and biological activities of anthocyanins. Phytochem, 2003, 64, 923–933.

Combs G.F., The Vitamins, Fundamental Aspects in Nutrition and Health. in 2nd Edition San Diego: CA:

Academic Press, 2001, 245-272.

Giroux H.J., De Grandpre G., Fustier P., Champagne C.P., St-Gelais D., Lacroix M., Britten M., Production and

characterization of Cheddar-type cheese enriched with green tea extract, Dairy Sci. Technol. 2013, 93, 241-254.

Han J., Britten M., St-Gelais D., Champagne C.P., Fustier P., Salmieri S., Lacroix M., Polyphenolic compounds

as functional ingredients in cheese, Food Chemistry, 2011, 124, 1589-1594. Izuagie A.A., Izuagie FO., Iodometric determination of ascorbic acid (vitamin C) in citrus fruits. Research

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Onion Cultivars, Czech Journal Food Science, 2013, 31, 283-291.

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tocopherol and anthocyanin extracts of blackberry, oil with synthetic alpha-tocopherol and anthocyanin extracts

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97

AUTHENTICITY OF DAIRY GOAT MILK EXISTING IN ROMANIAN

MARKET

Ramona Maria IANCU*

*PhD Fellow, SOP HRD/159/1.5/s/133675 Project, Lucian Blaga University from Sibiu, Partner, Romania

Abstract: In our days, many consumers are asking about the benefits of the authentic products and their

availability of consumption (Beverland, 2005), in a production world where almost all the products are mass-

produced. (Gilmore and Pine II, 2007) This study was made to perceive the consumers need for authentically

dairy goat milk product made in Romania, because the trend is to seek alternatives for authenticity. (Leigh et

all, 2006) This study identifies three characteristics of authenticity in our country for dairy goat milk

products: understanding the concept of authenticity, consumer’s knowledge on product authenticity analysis,

the new modalities concerning the counterfeiting of consumer protection regarding the dairy goat milk

products and the need to consume authentic goods by refusing to buy imitation products. Based on

questionnaires, one hundred consumers were interviewed during four month (January-April 2015), in

different markets and especially in our university, also looking to find the opinion of the young people. A

valuable and needed analyze to establish the authenticity of a specific dairy goat milk products using the

major milk proteins polymorphisms as genetic markers is well known for the researches in Romania as a

method of control, but unknown for the consumers.(Bâlteanu, 2010 ) This method could be helpful in taking

a decision to buy original and authentic dairy goat milk products.

Keywords: product, authenticity, consumer.

INTRODUCTION

In our days, many consumers are asking about the benefits of the authentic products

and their availability of consumption (Beverland, 2005), in a production world where almost

all the products are mass-produced. (Gilmore and Pine II, 2007) In meantime, an increasing

number of consumers wondering about what they consume and about how healthy they are.

Often, the decision of buying different products depends on price and how the product taste is

fulfilled them.(Goulding, 2000; Grayson and Martinec, 2004) For many years, consumers

buy to complete a specific level of their pleasure, making a continuous cycle of acquisition,

becoming in so many times dissatisfied. This study was made to perceive the consumers need

for authentically dairy goat milk product made in Romania, because the trend is to seek

alternatives for authenticity. (Leigh et all, 2006)



98


This study identifies three characteristics of authenticity in our country for dairy goat

milk products: understanding the concept of authenticity, consumer’s knowledge on product

authenticity analysis, the new modalities concerning the counterfeiting of consumer

protection regarding the dairy goat milk products and the need to consume authentic goods by

refusing to buy imitation products.

Based on questionnaires, one hundred consumers were interviewed during four month

(January-April 2015), in different markets and especially in our university, also looking to

find the opinion of the young people.


From the data that have been collected the interviewed consumers send a valuable

feed-back. The meaning of authenticity is based on the reality and truth, including terms such

as “ethical, natural, honest, simple, unspun, sustainable, beautiful, rooted and human”.

(Boyle, 2003) The results of this study are a link between Boyle description and Munoz

definition “original, genuine, unique, traditional and real”. (Munoz et all, 2006)

Also, the data interpretation proves the consumer’s concerns about their experiences

through their need and emotions, less about the scientific description of an authentic product.

A valuable and needed analyze to establish the authenticity of a specific dairy goat

milk products using the major milk proteins polymorphisms as genetic markers is well known

for the researches in Romania as a method of control, but unknown for the consumers.

(Bâlteanu, 2010) This method could be helpful in taking a decision to buy original and

authentic dairy goat milk products.(Bâlteanu et all, 2010)

In accordance to collected data, a statistical interpretation was made.

The regression model below is estimated linear regression model parameters and its

validity on a sample of 100 respondents. Data refer to the average time in seconds required to

complete the questionnaire (x) and the number of people who answer the 10 questions in the

questionnaire (y).

By the software EXCEL / data analysis / regression yielded the following data

parameter estimation linear regression model, the significance of the model parameters for a

5% significance level, the intensity between the two variables, model validity and possible

residual errors.



99

SUMMARY OUTPUT

Regression Statistics Multiple R 0,927 R Square 0,859328 Adjusted R Square 0,849227 Standard Error 1,941209 Observations 100

ANOVA

df SS MS F Significance

F Regression 1 2278,939 2278,939 604,7667 1,03E-43 Residual 99 373,0611 3,768294 Total 100 2652

Coefficients Standard

Error t Stat P-value Lower 95% Upper 95%

Intercept 0 #N/A #N/A #N/A #N/A #N/A 25 0,112053 0,004556 24,59201 5,91E-44 0,103012 0,121094

Figure 1. Representation of the number of respondents with positive responses



100

Figure 2. Chart showing the link between the number of positive responses and the average

time needed to complete the questionnaire for the 100 respondents

Figure 3. Distribution chart errors on estimated average time to complete the questionnaire

for the 100 respondents

Interpretation the results of Table SUMMARY OUTPUT

Multiple R = 0.927, indicates that the average time needed to complete the

questionnaire shows a strong link with the number of positive responses R Square = 0,859328

shows that 85% of the variation of positive responses is explained by the average time



101

required completing the questionnaire. Standard error = 1.941209 shows that not all points

are on regression.

Interpretation of the results in Table ANOVA

In order to validate the test regression model the test F is calculated. It shows the

value 604.7667 and the value of the Significance F = 1.03E-43, a value which is less than

0.05. These results establish that the regression model is valid. The term free, intercept is

where the variable factor is 0, so questionnaires will be 0 if the average time-on is 0. P-value

presents the value 5.91E-44.

CONCLUSIONS

Between the two variables X (average time in seconds required to complete the

questionnaire) and Y (the people who answer the 10 questions in the questionnaire) there is a

significant dependency.

The number of positive responses correlates with time of completion.

The regression model is valid and can be used to analyze dependence of the two variables

selected. P-value is significant because it is smaller than 0.05.

Since previous studies have pointed this consumer trend, this study identified and

generated more listings of authenticity from the point of view of consumers.

Our research found that consumers who need and want to buy authentic products

remain faithful to authentic attributes of the product, and, in the end, they reject the imitation

goods.

ACKNOWLEDGEMENTS

This paper is supported by the Sectoral Operational Programme Human Resources

Development (SOP HRD), financed from the European Social Fund and by the Romanian


REFERENCES Bâlteanu V.A., Vlaic A., Pop F.D., Carşai T.C., Pascal C., Creangă Şt., Zaharia N., Pădeanu I., Voia O.S., Sauer

M., Sauer I.W., The study of αS1-casein genetic marker polymorphism in Carpathian goat breed: synthesis of

2009 research (Project PN II 52104/2008). Lucrări Ştiinţifice Seria Zootehnie Iaşi, 53: 321-325, 2010



102

Bâlteanu V. A., The study of major milk proteins genetic polymorphisms in the main cattle, buffalo, sheep and

goat breeds from romania with the aim of using them as genetic markers in breeding and traceability, PhD

thesis, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Doctoral School Faculty of

Animal Husbandry and Biotechnology, Cluj Napoca, 2010

Beverland M., Crafting brand authenticity: The case of luxury wines, Journal of Management Studies 42(5),

1003-1029, 2005.

Boyle D., Authenticity: Brands, Fakes, Spin and the Lust for Real Life, London: Harper Perennial, 2003.

Gilmore J.H., and Pine II B.J., Authenticity: What Consumers Really Want, Boston: Harvard Business School

Press, 2007.

Goulding C., The commodification of the past, post-modern pastiche, and the search for authentic experiences at

contemporary heritage attractions, European Journal of Marketing 34(7), 835-853, 2000.

Grayson K. and Martinec R., Consumer perceptions of iconicity and indexicality and their influence on

assessments of authentic market offerings, Journal of Consumer Research 31, 296-312, 2004.

Leigh T W., Peters C. and Shelton J., The consumer quest for authenticity: The multiplicity of meanings within

the MG subculture of consumption, Academy of Marketing Science. Journal 34(4), 481-493, 2006.

Munoz C.L., Wood N.T., Solomon M.R., Real or blarney? A cross-cultural investigation of the perceived

authenticity of Irish pubs, Journal of Consumer Behaviour 5, 222-234, 2006.



103

COMPARISON BETWEEN TWO INSTRUMENTS FOR EVALUATION

OF DOUGH RHEOLOGY DURING KNEADING Mihai OGNEAN1, Olga DRĂGHICI1, Maria Lidia IANCU1

Faculty of Agricultural Sciences, Food Industry and Environmental Protection, “Lucian Blaga” University of

Sibiu, Victoriei Street, 10th, Sibiu, Romania

Abstract: Dough rheology is related with baking quality of wheat flour and empirical rheometry are widely

used by scientist and technologist in cereal processing. Different instruments are in use and new other are

designed. The new instruments need confirmations. In this work two instruments were compared an old,

classic, instrument, Farinograph with a new one, Flourgraph. The instruments were very similar in geometry

and working principle and work with the same procedure, delivery the same dough characteristic properties

derived from curve interpretation. Good correlations were observed for the instruments evaluated, the

regressions coefficients were above 0.9000. The highest regression coefficient was for flour’s capacity of

hydration, 0.9740, and the lowest for the dough stability, 0.9228. The results obtained on Flourgraph were

consistent and reproducible, with similar standard deviation and confidence interval.

Keywords: rheology, dough, Farinograph, Flourgraph E6

Introduction

The visco-elasto-plastic properties of dough are own to gluten from wheat flour

(Weipert, 2006). During the kneading the gliadins and glutenins from wheat flour are

hydrated and after that they combine to form a visco-elastic material named gluten which

could be separated by washing in a stream of water. Gluten forms in dough an extended

network which trapped inside the other components of dough as starch granule, bran

fragments, cells of yeast and bacteria and other (Bordei & Toma, 2000). Pellicle of gluten

covered starch granule and form a complex matrix with rheological properties similar to

gluten (Ognean, et al., 2005). This complex material forms pellicle able to retain gas as

bubbles during kneading defining the dough porosity. Gas bubbles expand during proving by

retaining carbon dioxide formed by yeast defining bread porosity after baking (Couvain,

2003). As a consequence of these the shape and volume of breads are close related to dough

rheology – properties of dough to be viscous and elastic and plastic. A balance of these

properties is necessary to form dough with good properties for baking. manhy scientist



104

consider that a very strong bond is among dough rheology and bread quality (Dobraszczyk,

2003). Rheology became important tool in the control of flour, evaluation of wheat potential

for baking and evaluation of different additives destined for dough conditioning.

Recently rheometer are more used for the measurement of dough rheology but empirical

instrument as Farinograph, Consistograph or Mixograph remain the most used instrument in

baking, milling, cereal commerce and even cereal science (Weipert, 2006).

Scientist designed different instrument to evaluate the dough rheology, through this

Farinograph was one of the most important, created for the study of dough behaviour during

mixing, one of the most critical phase in breadmaking. Others instruments were constructed.

Through the main used instruments are also Valorigraph, Mixograph, Consistograph and Do-

Corder. These instruments are made by different producers of lab instruments for cereal

industry and other instruments, more or less similar are produced now (Bloksma & Bushuk,

1988). Some improvements are added as the possibility to increase the temperature to

simulate the baking process as the case of Rapid Visco Analyser (Weipert, 2006). Other

instruments, as Flourgraph E6, just reproduce the design of other instruments and allow the

access to technology for lower price. These new instruments must offer similar information

as the original one. The aim of this work is to compare two instruments, the classic

Farinograph made by Brabender and Flourgraph E6, made by Haubelt. The Flourgraph E6

have a smaller mixing bowl, for 100 g of flour which allow the use of small quantity of flour

but still deliver enough quantity of dough for extensional test with Brabender’s Extensograph

or the similar Haubelt’s Flourgraph E7.

Despite new rheometer are widely used in food industry the old fashion empirical

rheometer remain very popular in cereal industry, delivering comprehensive information to

scientist and technologist in baking.


Flours testing was done in strict compliance with the methods Farinograph Method

for Flour, SR ISO 5530-1 / 199 equivalent with Method No. ICC 115/1.

We analysed the following parameters:

Maximum consistency (C expressed in UB - Brabender Units or UH - Haubelt Units);

Corrected hydration capacity for humidity and consistency;

Development time (DT in minutes);



105

Dough stability (S, in minutes);

Degree of softening (DS12, expressed in UB and UF);

Quality number (time required to lower the consistency of the dough with 30 units as

determined from reaching maximum consistency, multiplied by a multiplication factor

10) .

For testing were used 10 commercial or special flours chosen so as to be as diverse and cover

a wide possible range of values. Two or three tests were run using Brabender Farinograph

and 5 to 7 test with Flourgraph E6, for each flour. The results were statistically analysed

using Excel and SPSS 10. Confidence interval was calculated for 95%. It was analysed

standard deviation and confidence interval for each characteristic, for each flour and

instrument separately. For a better appreciation of the size of these parameters they were

expressed as the % of average analysed characteristics. Tests on the two instruments were

made within a maximum of 24 hours to prevent any change in the flours due to the natural

maturation process.

For tests were used Brabender Farinograph-E and Flourgraph E6.The Flourgraph E6

was kindly offered by Haubelt, Germany through Sartorom Romania.


Dough consistency

The hydration capacity was determined in a previous test, according with the method.

This value was used in the rheological test of flours. The same amount of water was used for

every test and the dough obtained had different consistencies. Average, maximum and

minimum consistencies are presented in Table 1. Although the amount of water remained

constant maximum dough consistency ranged deviation is different for each machine.

After data analysis was not observed big differences between the values obtained on both

instruments. For three flours the standard deviation and confidence interval was lower for

Flourgraph E6 while in 7 cases the values were lower for Farinograph

Corrected hydration capacity (CH14 )

CH14 represent the amount of water required to obtain a dough consistency of 500

units of 100 kg of flour with humidity equal to the reference humidity (14%). Corrected

hydration capacity is calculated by the software of the two instruments based on the

maximum value of the consistency of the dough, the amount of added water (hydration



106

capacity) and the sample humidity. Based on these data the instrument’s software make

corrections and calculates a new value that corresponds to dough consistency of 500 units and

made from flour with humidity of 14%. In the table no. 2 are presented the average,

maximum and minimum CH and standard deviation and confidence interval.

Table no 1. Statistical indicators for dough consistency

Sam

ple

Brabender Farinograph Haubelt Flourgraph E6

No

of

anal

ysis

C av

erag

e

Cm

ax

C m

in

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

No

evid

e0nc

e

C av

erag

e

Cm

ax

C m

in

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

F1 3 500 502 499 1.53 1. 7 6 502 506 497 3.74 3.0 F2 3 502 510 496 7.21 8.2 6 511 539 497 15.02 12.0 F3 3 501 505 499 3.46 3.9 7 496 505 485 6.90 5.1 F4 3 497 503 493 5.13 5.8 6 508 511 506 1.72 1.4 F5 3 493 497 490 3.51 4.0 5 501 506 498 3.56 3.1 F6 2 504 505 502 2.12 2.9 6 505 510 500 3.31 2.6 F7 3 507 515 502 6.81 7.7 6 507 511 499 4.27 3.4 F8 3 497 502 488 7.57 8.6 7 501 508 489 6.47 4.8 F9 3 500 504 493 5.86 6.6 6 506 522 496 9.23 7.4 F10 3 496 499 493 3.06 3.5 5 496 503 490 5.63 4.9

Table no 2. Statistical indicators of corrected capacity of hydration

Sam

ple


aver

age

CH

14

CH

14

max

CH

14

min

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

,

aver

age

CH

14

CH

14 m

ax

CH

14

min

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

F1 58.2 58.3 58.2 0.06 0.1 58.2 58.3 58.1 0.08 0.1 F2 59.8 60 59.7 0.15 0.2 58.7 59.3 58.3 0.35 0.3 F3 58.6 58.7 58.6 0.06 0.1 58.4 58.6 58.2 0.15 0.1 F4 62.8 63 62.7 0.15 0.2 62.1 62.2 62.0 0.06 0.1 F5 54.1 54.3 54 0.15 0.2 52.0 52.1 52.0 0.05 0.0 F6 53.1 53.1 53 0.07 0.1 50.5 50.6 50.3 0.10 0.1 F7 65.0 65.2 64.9 0.15 0.2 63.3 63.4 63.1 0.10 0.1 F8 60.9 61 60.7 0.15 0.2 60.2 60.4 59.9 0.16 0.1 F9 58.7 58.8 58.5 0.17 0.2 55.8 56.1 55.6 0.22 0.2 F10 58.3 58.4 58.2 0.10 0.1 56.4 56.6 56.3 0.13 0.1

In terms of statistical dispersion of data it is very similar for the two instruments. The

values determined on Flourgraph E6 were slightly higher than the values obtain on Brabender

Farinograph. In figure 1 are presented graphically the CH’s values obtained on the two

instruments. The correlation between the instruments is very high, R2 was 0.9740. CH varied



107

between 50.1 and 65.0%. The mathematic equation which describe the linear relation

between the two sets of values was y = 1.106 ∙ x – 7.632.

Figure 1. Recorded corrected CH determined on Flourgraph E6 vs. Farinograph

Development time (DT)

This parameter provides information about the time necessary for a complete

development of dough during mixing.

In statistical analysis of the values we renounced at flour F6, which were a strong

flour and the curves obtained on the instruments were very different. Curve obtained on

Brabender Farinograph curve shows a single peak while the instrument manufactured by

Haubelt showed a second maximum, greater than the first and the methodology considered

this one as the real peak. The results are presented in Table No. 3.

Table no. 3 Statistical indicators for dough development time (DT)

Sam

ple


DT

DT

Max

DT

min

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

DT

DT

Max

DT

min

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

F1 2 2.3 2 0.15 2 2.1 2.5 1.9 0.23 0.2 F2 2 1.7 1.5 0.12 2 2.0 2.4 1.6 0.29 0.2 F3 2 2.2 2 0.12 2 2.1 2.5 1.8 0.22 0.2 F4 6 6.7 4.8 0.96 6 5.7 6.1 5.2 0.36 0.3 F5 4 4 3.7 0.15 4 4.2 4.5 3.8 0.25 0.2 F7 6 5.7 5.5 0.12 6 4.6 5.0 4.3 0.24 0.2 F8 2 1.7 1.7 0.00 2 1.8 1.9 1.6 0.11 0.1 F9 2 2 1.9 0.06 2 2.3 2.9 2.0 0.36 0.3 F10 2 2.4 1.8 0.31 2 2.2 2.2 2.1 0.04 0.0

CH14 Brabender

7068666462605856545250

CH

14 H

aube

lt

70

68

66

64

62

60

58

56

54

52

50

F10F9

F8

F7

F6

F5

F4

F3 F2F1



108

The linear correlation between the sets of values obtained through different

instruments was R 2 = 0.9468, and for this kind of instruments indicated a good correlation.

Mathematical equation which describe the relations between the values of development time

was y = 0.8393 ∙ x + 0.4914. The multiplication factor was close to 1 but different, which

indicate differences between the instruments and the behaviour of flours during mixing.

Stability (S)

Dough stability provides information about how much time the dough consistency

remains high. It determined that the period in which the upper edge of the curve is above the

line that defines the maximum consistency of the dough. In Table No 4 are presented the

statistical indicators of stability. Flour F6 was excluded again.

Table no. 4. Statistical indicators of stability dough

Sam

ple


S av

erag

e

S m

ax

S m

in

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

S av

erag

e

S m

ax

S m

in

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

F1 2 2.3 2 0.15 2 2.1 2.5 1.9 0.23 0.2 F2 2 1.7 1.5 0.12 2 2.0 2.4 1.6 0.29 0.2 F3 2 2.2 2 0.12 2 2.1 2.5 1.8 0.22 0.2 F4 6 6.7 4.8 0.96 6 5.7 6.1 5.2 0.36 0.3 F5 4 4 3.7 0.15 4 4.2 4.5 3.8 0.25 0.2 F7 6 5.7 5.5 0.12 6 4.6 5.0 4.3 0.24 0.2 F8 2 1.7 1.7 0.00 2 1.8 1.9 1.6 0.11 0.1 F9 2 2 1.9 0.06 2 2.3 2.9 2.0 0.36 0.3 F10 2 2.4 1.8 0.31 2 2.2 2.2 2.1 0.04 0.0

Figure 2. Correlations between dough stability determined on Farinograph and Flourgraph

The correlation between the instruments was poorer than for development time and also the

multiplication factor in the equation.

y = 0.828x + 0.1286R2 = 0.9228

0.01.02.03.04.05.06.07.08.09.0

10.0

0 2 4 6 8 10Brabender S

Hau

belt

S



109

Dough softening

Statistical indicators of the values obtained for dough softening are presented in table

5. The values are similar for the instruments which were compared. The coefficient of

regressions was good, 0.9288. Distribution of values obtained through Flourgraph was better

than in the case of Farinograph, the values were closer to average value. The mathematical

equation which describe relation between softening measured on Flourgraph and dough

softening measured on Farinograph was y = 0.8022 ∙ x + 8.9327.

Quality number (QN)

This number is closely related to the time required to decrease doughy consistency by

30 units from maximum. The higher it is the number the stronger it is the flour.

The values obtained for the quality number were distributed over a larger interval

around the mean value, for both instruments. In the case of Flourgraph the values were more

disperse than for Farinograph. A good correlation was observed between the instruments, R2

= 0.9474 but the importance of other factors was great. The constant from the mathematical

equation which describe the relation between the quality numbers measured on Flourgraph

and Farinograph was 3.911. The equation was y = 0.7723 ∙ x + 3.9111. The Flourgraph

delivered lower values for the quality number than Farinograph.

Table No 5. Statistical indicators of dough softening

Sam

ple


Ave

rage

D

S 12

DS 1

2 m

ax

DS1

2 m

in

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

Ave

rage

D

S 12

DS 1

2 m

ax

DS 1

2 m

in

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

F1 48 50 47 1.53 1.7 36 45 29 5.96 4.8 F2 112 120 106 7.21 8.2 98 108 88 7.78 6.2 F3 46 47 45 1.00 1.1 38 47 30 6.16 4.6 F4 20 22 16 3.21 3.6 19 20 15 1.86 1.5 F5 51 53 50 1.53 1.7 38 43 33 3.58 3.1 F6 F7 35 37 34 1.73 2.0 41 45 36 3.08 2.5 F8 87 93 78 8.14 9.2 71 75 62 4.35 3.2 F9 58 61 55 3.00 3.4 51 61 41 7.65 6.1 F10 61 64 57 3.61 4.1 53 60 47 4.95 4.3



110

Tabel No 6. Statistical indicators of the quality number Sa

mpl

e

Brabender Farinograph Haubelt Flourgraph E6 Q

N av

erag

e

QN

max

QN

min

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

QN

ave

rage

QN

max

QN

min

Stan

dard

de

viat

ion

Con

fiden

ce

inte

rval

F1 50 51 47 2.31 2.6 41 58 34 8.68 6.9 F2 27 29 26 1.53 1.7 29 34 23 4.54 3.6 F3 65 80 49 15.52 17.6 44 89 24 23.29 17.3 F4 119 123 116 3.61 4.1 102 110 86 8.86 7.1 F5 72 75 71 2.31 2.6 66 73 61 4.55 4.0 F6 F7 93 95 91 2.08 2.4 69 78 64 5.97 4.8 F8 30 34 27 3.51 4.0 27 29 25 1.27 0.9 F9 39 40 38 1.00 1.1 37 50 29 8.50 6.8 F10 41 43 38 2.52 2.8 35 40 28 4.76 4.2

Conclusions

The new rheometer Flourgraph E6 was very similar to the Farinograph-E in geometry,

working procedure and results. Good correlations were observed among the values of

characteristics obtained through instruments. The regression coefficients were above 0,9000.

The strongest correlation was observed in the case of flour’s capacity of hydration while the

weakest correlation was observed for quality number. For the quality number we observed

the highest standard deviations and confidence interval for the both instruments. For the

majority of flours Farinograph the values obtained for different characteristics showed lower

standard deviation. Differences were observed for flour F6 – the curves obtained using both

instruments were different in shape, Flourgraph reveal a second peak.

Differences among instruments could be explained by decalibration or intrinsic

differences of instruments. References.

Bloksma, A. H. & Bushuk, W., 1988. Rheology and chemistry of dough. În: Y. Pomeranz, ed. Wheat:

Chemistry and Technology, 3rd. St. Paul, Mn, USA: American Association of Cereal Chemists, pp. 131-218.

Bordei, D. T. F. & Toma, M., 2000. Știința și tehnologia panificației. București: Ed. Agir.

Couvain, S. P., 2003. Breadmaking: an overview. In: S. P. Couvain, ed. Bread making. Improving quality.

s.l.:Woodhead Publishing.

Dobraszczyk, B. J., 2003. Measuring the rheological properties of dough. In: S. P. Couvain, ed. Breadmaking.

Improving Quality. s.l.:Woodhead Publishing.

Ognean, M., I, D. & V, G., 2005. Procese biotehnologice în panificaţie,. Sibiu: Ed. Universităţii „Lucian Blaga”

din Sibiu.

Weipert, D., 2006. Rheology. In: Future of Flour - A compendium of Flour Improvement. s.l.:Agrimedia.



111

RHEOLOGY OF WAXY MAIZE STARCHES AS TOOL FOR

INDUSTRIAL APPLICATIONS

Elena-Roxana Tufeanu, ŞtefanOancea, Monica Mironescu, Ion Mironescu, OvidiuTiţa

Faculty of Agricultural Sciences, Food Industry and Environmental Protection, “Lucian

Blaga” University of Sibiu, Victoriei Street, 10th, Sibiu, Romania

Abstract: The paper investigates the rheological behaviour of three types of waxy maize starch: native,

pregelatinizated and acetylated starches in order to find the appropriate application for each starch type as

function of temperature, from room to gelatinisation temperature. A rotational rheoviscosimeter with

geometry with concentric cylinders is used. The results indicate the pregelatinised starch as the most

appropriate for uses to increase viscosity in instant foods obtained at room temperatures and in warm water

(50oC), whereas acetylated starch is very suitable for increasing viscosity in products treated at higher

temperatures (80oC). native waxy maize starch is recommended for products which doesn’t require high

viscosities.

Key words: waxy maize starch, rheology, viscosity, temperature

Introduction

Starch is one of the widespread natural products and a main component of foods

(Song, 2013; Nemtanu, 2008). The remarkable physicochemical properties of this

carbohydrate are due to amylose and amylopectin, two polymeric components (Wang, 2012).

Normal starch contains amylose between 20 and 30%, w/w, high-amylose starch has in its

composition amylose content >50%, w/w and waxy starch contains only traces of amylose

(Achayuthakan&Suphantharika, 2008; Preiss, 1991).

Use of starch in the food industry is limited due to tendency towards retrogradation,

low shear resistance and thermal stability. To improve such properties is recourse to

modifystarch by various treatments: physical, chemical, enzymatic or combined(Wurzburg,

2006; Abbas et al. 2010).Modifications allow to maintain texture despite stress at processing,

enhance product aesthetics, reduce production costs, ensure constantquality product, and

extend shelf life (Banu, 2009; Be Miller, 2009; Berski, 2011). Also modified starch have

been developed in order to improve water holding capacity, freeze-thaw and low pH



112

resistance as to reinforce its binding properties and minimize syneresis in food (Abbas et al.

2010).

Possible application of starch as thickeners or as gelling agents are determined by

rheological properties. Viscosity is the basic rheological characteristic of starch paste/gel.

Because therheological behaviourofstarchpastes/gelsis veryimportantintheprocessing,

theirrheologicalpropertiesare investigatedquite intensively. They depends on many factors as

starch type, starch concentration, temperature or shearing rate (Mironescu et al., 2011).

This paper investigates the evolution of the viscosity of various types of maize starch

by the action of temperature and mechanical stress, in order to identify the most suitable

practical applications for different waxy maize starch types, including native, pregelatinised

and acetylated starch.


Three types of waxy maize starch were used, all purchased from the Romanian

commercial system:

- Native starches (from two sources, abbreviated with N1, N2);

- Physically modified starch, namely pregelatinised starches (from two sources,

abbreviated with P1, P2);

- Chemically modified starches, namely acetylated di-starch adipate E1422

(abbreviated with M1 and M2).

5 % aqueous suspensions from all samples were used.

The rheological measurements were performed by using a HAAKE VT® 550

rotational rheoviscosimeter having a geometry with concentric cylinders MV DIN connected

to a water bath. The measurements were carried out at 25oC, 50oC and 80oC at speed range

from 0 to 500 rpm / s in 10 steps. The obtained data were analysed with the RheoWin v.3.5

software.


As figure 1a shows, pregelatinised starches P1 and P2 show pseudoplastic behaviour;

due to the physical changes that have been subjected, the pregelatinised starch has higher

values of viscosity at low temperatures, the result indicating them as suitable for products

which are manufactured at low temperatures. In this way, pregelatinised starch P2 is the most



113

suitable for applications at room temperatures in instant foods as cold creams or sauces. For

this starch type, very low shear rates are required to obtain high viscosity.

Even P2 is also a pregelatinisedstarch, rheological analysis indicates that viscosity is

not so strong increasing as in the case of P1. This result shows the importance of the

pregelatinisation treatment, as evidenced by Mironescu et al. (2006).

At 25oC, viscosities of native and chemically modified starches attire values of close

to zero (figure 1b) and they have quasi-newtonian behaviour with a dilatant tendency.

Figure 1a. Starch

viscosity curves at

25oC

Figure 1b. Detail:

viscosity curves of

native and chemical

modified starch at

25oC

Figures2a and 2b show the evolution of viscosity as function of the shear rate for

samples at 50oC. For all starch types, no big differences are observed compared with the

analysis at room temperature, indicating that no changes in the starch structure (meaning

gelatinisation) starts at this temperature.

Starch sample P2 is the most suitable for applications in instant foods obtained by

adding warm water where higher viscosity is required as puddings, warm creams or warm

sauces.



114

Viscosity of the pregelatinised starch P2 evolves very interesting from 25oC to 80oC.

As the rheological analysis at 25oC (figure 1a), 50oC (figure 2a) and 80oC (figure 3a) shows,

no changes are induced by temperatures for P2, meaning that gelatinisation is already

finished at the obtaining of this type of pregelatinised starch. More, the increase of

temperature give products with smaller viscosities.

Figure 2a. Starch

viscosity curves at

50oC

Figure 2b. Detail:

viscosity curves of

native and chemical

modified starch at

50oC

The other pregelatinised starch analysed, P1, behaves similarly, but with much lower

viscosity as P1 and viscosity decreases with temperature increase, as figures 1a, 2a and 3a

show.

At 80oC the rheological behaviour changes for the chemicallymodified starch M1

which become high viscous(figure 3a), making him suitable on technological applications

where very high viscosities are necessary. Modified starch M1 is highly pseudoplastic at high

temperatures this behaviour being due to gelatinisation. Sample M1 has the highest viscosity,

followed by N2 which become also pseudoplastic at high temperatures.



115

M2 continues to have very low values of viscosity at 80oC; itsrheological behaviour

remains quasi–newtonian in a large temperatures range, from 25 to 80oC. The analysis

indicates M2 as the most adequate for easy transport through conducts even at high

temperatures.

Viscosity of P1 remainsat very low levels, indicating the native starches also as

adequate for easy transport through conducts.

Figure 3a. Starch

viscosity curves at

80oC

Figure 3b. Detail:

viscosity curves of

someof the analysed

starches at 80oC

Conclusions

Rheological analysis is very important for finding valuable applications for starches.

Native starches are useful in products in which no high viscosity increase is necessary even at

high temperatures. Pregelatinisedstarch are recommended for instant foods obtained with

water at room temperature to increase their viscosity, whereas high viscosity increase is

obtained with acetylated di-starch adipate at 80oC.



116

Even the name of the modified starch is the same, the preliminary treatment is very

important.

References 1. Abbas, K. A., Khalil, S. K., &Hussin, A. S. M., Modified starches and their usages in selected food

products: a review study. Journal of Agricultural Science, 2010, 2(2), 90–100.

2. Achayuthakan, P., &Suphantharika,M., Pasting and rheological properties of waxy corn starch as affected

by guar gum and xanthan gum. Carbohydrate Polymers, 2008, 71(1), 9–17.

3. Banu C., Industriaamidonuluişi a produselorderivate, Tratatde industriealimentară, Ed. ASAB, Bucureşti,

2009.

4. Bao Wang, Li-jun Wang, Dong Li, Qing Weia, BenuAdhikaric, The rheological behavior of native and

high-pressure homogenized waxy maize starch pastes, Carbohydrate Polymers, 88, 2012, 481– 489.

5. BeMiller, J. N., & Whistler, R. Starch — chemistry and technology, 2009. (3rd ed.).

6. Berski W., Ptaszek A., Ptaszek P., Ziobro R., Kowalski G., Grzesik M., Achremowicz B., Pasting and

rheological properties of oat starch and its derivatives, Carbohydrate Polymers 83, 2011, 665–671.

7. Chang-Geun Song · Moo-YeolBaik, Byung-Yong Kim, Rheological Properties of Native Maize, Waxy

Maize, and Acetylated Maize Starches, and Applications in the Development of Food Products. J Korean

SocApplBiolChem, 2013, 56, 63−68

8. Mironescu M., Iancu M.L., Mironescu V., Quality assurance at the obtaining of pregelatinised starches,

Annales of the University of Craiova, Secţiunile “Biologie, Horticultura,

TehnologiaPrelucrăriiProduselorAgricole, IngineriaMediului”, 2006, ISSN 1435-1275, vol. 11, p. 133-138

9. Mironescu M., MironescuI.D., Oprean L., Steady and oscillatory shear behaviour of semi-concentrated

starch suspensions, Procedia Food Science, 2011, 1, 322-327

10. Nemtanu M.R., Brasoveanu M., Aspects regarding the rheological behavior of the wheat starch treated

with accelerated electron beam, Rom. Journ. Phys., Vol. 55, 2010, Nos. 1–2, P. 111–117, Bucharest.

11. Preiss, J., Biology and molecular biology of starch synthesis and its regulation. Oxford Surveys of Plant

Molecular and Cell Biology, 7, 1991, 59–114. 12. Wurzburg, O. B., Modified starches. In A. M. Stephen, G. O. Phillips, & P. A. Williams (Eds.), Food

polysaccharides and their applications (2nd ed., pp. 87–118). Florida: CRC, 2006.



117

ADJUSTMENT OF CORRUGATIONS NUMBER/CM, FOR THE BREAKING

ROLLERS, RELATED TO THE GRINDING RESISTANCE

OF THE WHEAT GRAIN

Cristina Anca DANCIU*

*The Faculty of Agricultural Sciences, Food Industry and Environmental Protection, Lucian Blaga University,

5-7 Ion Ratiu str, Sibiu, ROMANIA,[email protected], http://saiapm.ulbsibiu.ro

Abstract: The heart of a flour mill is the mill stand rolls. These corrugated rolls are essential in the break

system to separate the endosperm from the bran and germ. The aim of the research is to identify the optimum

technological parameters regarding the roll corrugation number per cm, for obtaining the maximum

technological efficiency, with minimum energy consumption and grinding resistance of the wheat grains, on

the first breaking step in the wheat milling process. The aim of the research is to study the influence on the

number of corrugations per cm circumference of grinding rolls on grinding resistance. The greater the

number of corrugations the circumference of the two rollers will be greater the shredding because it

increases the number of shear and compression zones of the particles between the two rollers , the particles

traveling over the grinding zone .

Keywords: wheat, grinding resistance, roll corrugations, energy consumption.

INTRODUCTION

Wheat is the largest crop produced for human consumption in the world. Wheat is

milled to remove the bran and germ and reduce the wheat kernel to flour to be used in various

baked and nonbaked goods (Bass, 1988).

In wheat flour milling the main grinding tool is the roller mill, in which the feed

material is passed between two counter-rotating rolls of, usually 250 mm diameter and with

either corrugated or smooth finish (Posner, Hibbs, 2005). The function of the break rolls is to

open the kernel and remove the endosperm from the bran layers; size the endosperm pieces.

In the conventional 5 break grinding system the first 3 breaks are designed to open the wheat

kernel, to break off the bulk of the endosperm and to size the endosperm properly; the last 2

breaks scrape the remaining endosperm off the bran without cutting up the bran. Depending

on the passage, cylinders may have corrugations cut in them for shearing action. Other



118

passages will use smooth surfaces for more of a compression grind. The number and shape of

these corrugations involve complicated technical points. The numbers range from 4 to 12

(exceptionally 14) per cm of the roll circumference (Danciu,2003) for the conventional 5

break grinding system. Still further to improve the separations the corrugations are cut at

varying angles in relation to the rolls' axes.

In Table1 are given values for corr. no./cm for the breaking rolls of a 75 t / 24

hours mill capacity.

Table 1 Roll parameters for different break steps

No. Break No. corrugation/cm (R)

Inclination %

/0 Roll disposition

1 B I 4 6 30/60 S/S 2 B IIM 5 8 30/60 S/S 3 B IIm 6 8 30/60 S/S 4 B IIIM 7 8 30/65 S/S 5 B IIIm 7,5(8) 8 35/65 S/S 6 B IVM 8 10 35/70 D/D 7 B IVm 8,5(9) 10 35/70 D/D 8 B V 10 12 40/70 D/D 9 B IV 11 12 40/70 D/D

10 M6 12 12 40/70 D/D 11 M7 12 12 40/70 D/D 12 M8 13 14 40/70 D/D

The corrugation (R) number per cm, located on fast roller passing during scroll by

a particle grinding area through a point at a speed equal to that of the particle, is :

RpRt nvvR .

vR – speed of the fast roll;

vp – speed of the particle in the breaking area;

- the period of time for particle to pass through the breaking area;

nR – corrugations number per cm

pRp

LRp

vvvvv

LvL

0

2

v0p – speed of the fast roll related to the particle;



119

11

11

22

KKLnR

LnKvKvR

Lnvvvv

vvLnvvR

Rt

RL

Lt

RLR

LR

LRR

LRt

K – the speed ratio for the fast and slow rolls.

The corrugations number per cm circumference of the roller depends on the

grinding path length,on the ratio of peripheral speeds of the two rollers.

The total number of corrugations acting on the grinding products will be equal to

the number of corrugations located on the corresponding grinding length of the roller plus the

number of corrugations from the slow roller to enter into interaction with the particles

crossing the breaking zone , plus the number of corrugations on the length of the

corresponding fast grinding roller which are added quickly to the corrugations of the fast

roller coming into interaction with the particles during crossing the breaking zone (Osborne

B.G., 2001).

Rp

RRRRRR

Lp

LLLLLL

nvLvLnnvLnR

nvLvLnnvLnR

.

.

LR RRR

but: RL nn

p

R

p

LR v

vvvLnR 2

and: Kvv

L

R

p

L

p

LR v

KvvvLnR 2

The total number of corrugations participating in the grinding area depends on the

length of the breaking zone , the number of corrugations per centimeter circumference of the

roller , the mean particle velocity in the area of breaking and the peripheral speed of the two

rollers.



120

Consequently, as the corrugations number will be higher per centimeter

circumference of roller , other parameters being equal , the more intense breaking process it

will be (Osborne B.G. et al, 2006).

An additional argument is that for the same distance between the corrugations peaks

() of the two rollers , with the increasing number of corrugations per cm, depth (H)

decreases also decreasing the maximum size of particles which can hide in the valley between

two consecutive corrugations, Fig. 1.

Fig. 1 Changes in the intensity of the grinding according to the number of corrugations / cm

21

22

11

21

aaHaHa

HH


The research influence on the number of corrugations per cm circumference of

grinding rolls on grinding resistance, was made by using 30 g samples Pegasus durum wheat

(73 % vitreousness) , soft wheat Dropia (27 % vitreousness) respectively , which were

subjected to grinding with facility for measuring resistance grinding cereals. The preparation

of the samples colected carried out according to the chess-board pattern method, after

cleaning with an Sadkiewicz Instruments Scourer.The physicochemical characteristics of the

wheat were evaluated as follows: the moisture content using the SR ISO 712 : 2005; the wet

gluten content, protein content using the NIR technique (Inframatic, model 8600, Perten



121

Instruments AB); vitreous kernel using the STAS 6283-2/1984 (farinotom apparatus,Pohl). It

was used a micromill designed for simultaneously measurements of the compression efforts

and the shearing efforts, in the same conditions as in the milling industry.The micromill is

equipped with rolls measuring 50 mm in length and 90 mm in diameter, 0,5 mm roll gap for

the first breaking step, roll disposition sharp-sharp (S/S), 2,5:1 differential speed ratio, 8%

inclination and 30/600 (α/β) profile. The study was conducted on 7 corrugations/cm and then

9 corrugations/cm.

The appreciation of the grinding resistance is made by measurements of the resistant

moment of the wheat grain, between the rollers, in the breaking process. The resistant

moment of the particles grounded between the rollers, is measured by a tensometric cell,

conected to a PC computer and managed with a software program.

RESULTS AND DISSCUSIONS

Results timing depending resistant wheat varieties are summarized in Fig.2 and Fig.

3 for 7corrugations / cm respectively 9 corrugations / cm . So when using 7 corrugations / cm

and for 9 corrugations / cm , Pegasus durum wheat variety has the highest resistant to

breaking moment .

Fig. 2 Variation of the resistant moment for 7 corrugations/cm

0

0,5

1

1,5

2

2,5

3

1 2 3 4 5 6 7 8No. of wheat grains

Res

ista

nt M

omen

t, Nm

PEGASUSDROPIA



122

Fig. 3 Variation of the resistant moment for 9corrugations/cm

Regarding the moment of resistance , in the grinding process, it is inversely

proprotional to corrugations number per cm ; as the no. corrug. / cm increases the shredding

is more intense when the grains of wheat resistant to crumbling is less, so for Pegasus variety

of durum wheat and soft wheat variety Dropia .

Figure 4 and Figure 5 according to the graphs show that optimizing energy

consumption in grinding grain and is based on no. corrug. / cm circumference roller . The

higher the number the greater the intensity shred rifluri increases and decreases power

consumption .

CONCLUSIONS

Number of corrugations per centimeter circumference of roller , directly affects

shredding . The greater the number of corrugations the circumference of the two rollers will

be greater the shredding because it increases the number of shear and compression zones of

the particles between the two rollers , the particles traveling over the grinding zone . Stepping

shred resistant cereals involves decreasing the time and also to reduce energy consumption in

the process of crumbling .

0

0,5

1

1,5

2

2,5

3

3,5


Res

ista

nt M

omen

t, N

m

PEGASUSDROPIA



123

Choosing the optimal number of corrugations / cm circumference grinding roll is

consistent with the technological stage of the shredding : breaking , meal , or meal grinding,

closely related to the type and quality of their products once the proportion of coatings and

endosperm .

Fig. 4 Moment resistance variation for the variety Pegasus

0

0,5

1

1,5

2

2,5

3

3,5

1 2 3 4 5 6 7 8

No. of wheat grains

Res

ista

nt M

omen

t, Nm

R7 R9



124

Fig. 5 Moment resistance variation for the variety Dropia

REFERENCES Bass, EJ, Wheat flour milling, In Y Pomeranz, Wheat: Chemistry and Technology, vol. 2, 3rd edition, pp. 1-68,

St. Paul Minnesota: American Association of Cereal Chemists , 1988.

Costin, I.:, Cartea morarului, Editura Tehnică, Bucureşti, 1988.

Danciu, I., Măcinarea cerealelor, Ed. Univ. Lucian Blaga Sibiu, 2000.

Osborne, B. G, Wheat Flour Milling. Part 2. In Cereals and cereal product:chemistry and technology, Aspen

Publishers Inc., Maryland, USA., 2001.

Osborne, B.G., Henry, R., Southan, M.D, Assessment of commercial milling performance of hard wheat by

measurement of the rheological properties of whole grain, 2006Journal of Cereal Science.

Posner,ES, Hibbs, AN, Wheat flour milling, St. Paul Minnesota: American Association of Cereal Chemists,

2005.

0

0,5

1

1,5

2

2,5

3

3,5

4


Res

ista

nt M

omen

t Nm

R7R9



125

IDENTIFICATION AND CHARACTERIZATION OF USEFUL

SUBPRODUCTS OF GRAPEWINE PRODUCTS,

CASE STUDY: WINE YEAST

Mihaela BALTES*

*PhD Fellow, SOP HRD/159/1.5/S/133675 Project, Romanian Academy Iasi Branch or “Lucian Blaga” University from Sibiu, Partner, e-mail: [email protected]

Abstract Yeast nitrogenous compounds represent an important factor when it comes to selecting them for industrial purposes, with a view to value them at a superior potential. The present paper aims at evaluating an important component of Saccharomyces cerevisiae industrial yeasts, i.e. the protein concentration. The samples were selected from four recognized Wine Centers: Apold, Blaj, Miercurea and Sebeș. We employed the Kjeldahl method. The results obtained showed that the samples from the Miercurea M2 Wine Center recorded a maximum value of 53.875 grams protein / 100 grams dry matter, while the minimum value was ascertained in the samples from the Sebeș Wine Center, S2 recording a value of 44.500 grams protein / 100 grams dry matter. The results obtained led to the recommendation that residual yeasts in the Miercurea Wine Center are used for industrial purposes, as they have an important protein concentration. Keywords: Saccharomyces cerevisiae wine yeasts, proteins, total nitrogen

INTRODUCTION

Wine yeast is an important grapewine subproduct. Wine yeast has a complex composition,

rich in nitrogenous substances, carbohydrates, vitamins, lipids and other useful compounds.

Regarding nitrogenous substances, the crude protein has the highest share, mainly made up of

amino acids and assimilable nitrogen fractions (Albers et al. 1996), (Oprean et al. 2012), (Tița

et al. 2008). Nitrogen is assimilated by graprevine and yeasts from various sources, such as

soil, water, external sources (Bell et al. 2005), (Gardner et al. 2002). Studies conducted until

now have shown that adding nitrogen in wine-making processes causes considerable effects

on the grapes' sensory palette, implicitly on wine, leading to an increase in the

biotechnological potential of yeasts involved in fermentative processes (Mendez-Costabel et

al. 2014), (Tița et al. 2011), (Rapp et al. 1995). Fermentation kinetics is more active,

formation of aroma compounds amplifies, and therefore the value of assimilable nitrogen also

rises in the yeasts involved in these processes. Another important aspect is the way in which

soils are irrigated and the concentrations of nitrogen based fertilizers used in their



126

fertilization. An optimum concentration leads to a balanced accumulation of nitrogen

compounds in yeasts, implicitly to the formation of the wine sensory palette. Other studies

have demonstrated that different strains of Saccharomyces cerevisiae yeasts present different

properties of assimilating nitrogen compounds. Cultivated in environments with various

concentrations of nitrogen compounds, they presented different capacities to assimilate them,

producing biomass in different quantities (Torrea et al. 2011), (Varela et al. 2004), (Ribereau-

Gayon et al. 2006).


Four Wine Centers were selected for the present study: Apold, Blaj, Miercurea și Sebeș.

Sixteen samples of white wine yeast were taken from these centers, marked according to their

origin with the initial of the Center and the sample order number: A1, A2, A3, A4, B1, B2,

B3, B4, M1, M2, M3, M4, S1, S2, S3, S4)

The yeast samples were dried, then triturated in the ball mill, so that the extraction of nitrogen

compounds was done at maximum potential. To determine the proteins, the Kjeldahl method

was employed. This method consists of mineralizing the yeast samples with sulfuric acid;

protein nitrogen is transformed in ammonium ions subsequently determined through titration

with a strong acid. When digestion is over, the solution in the mineralization flask is

alkalinized by adding NaOH. The gaseous ammonia formed is captured in a collector flask

containing a boric acid solution. The ammonia is retained in solution as salt; the content of

nitrogen is determined through titration of the ammonium borate formed with a titrimetric

solution of sulfuric acid. The volume of acid solution necessary to titrate is equivalent to the

quantity of ammonia released. It is expressed in g/100g dry matter; the crude protein is given

by the quantity of total nitrogen identified through the Kjeldahl method X 6.25.

RESULTS AND DISCUSSIONS

As shown in Figure 1, the amount of total nitrogen determined through the Kjeldahl method

in the yeasts from the Apold Wine Center fall between 7.24 g/100g dry matter and a

maximum of 8.11 g/100g dry matter.



127

Figure 1. The amount of total nitrogen determined in the yeasts from Apold Wine Center

expressed in grams/100 grams dry matter

Samples A1 and A3 present the lowest nitrogen concentrations, around 7.3, while samples A2

and A4 present concentrations close to 8.

According to Figure 2, the yeasts from Blaj Wine Center have a near constant nitrogen

concentration, the values recorded being close to 8. The differences ascertained are of the

order of hundredths, which proves that the strains used in the fermentative processes come

from the same lot or producer, having no distinctive notes. Thus, we observe a maximum

value of 8.23 in sample B1 and a minimum value of 7.92 in sample B2; the percentage

difference between the two values is 3.8%.

Figure 2. The amount of total nitrogen determined in the yeasts from Blaj Wine Center




128

Miercurea Wine Center is a special case, as it has yeasts of oscilating nitrogen

concentrations; the samples differ with approximately 12%. Thus, according to Figure 3, it is

ascertained that the minimum value detected is of 7.55 grams/100 grams dry matter in sample

M3, and the maximum value reaches 8.62 grams/100 grams dry matter in sample M2. This

confirms the fact that, in the grape processing system, they used culture yeasts having

different biotechnological properties.

Figure 3. The amount of total nitrogen determined in the yeasts from Miercurea Wine Center


Figure 4. The amount of total nitrogen determined in the yeasts from Sebeș Wine Center




129

In the Sebeș Wine Center, we ascertain that the yeasts employed in the alcoholic fermentation

have come from different producers: the values recorded are low, about 7-8 grams/ 100

grams dry matter, yeasts having classic biotechnological properties, not presenting

spectacular accumulations of nitrogenous substances. In this context, they are a bit different

from the other samples; the values recorded are shown in Figure 4.

Following the calculations made to determine crude protein (Figure 5), we can say that

samples M2 from the Miercurea Wine Center have a maximum value of 53.875 grams / 100

grams dry matter, while the minimum value is recorded in the samples from the Sebeș Wine

Center, S2 recording a value of 44.500 grams / 100 grams dry matter.

Figure 5. The amount of crude protein calculated in the yeasts from Sebeș Wine Center


CONCLUSIONS

The wine yeasts selected to be studied have presented different protein values, depending on

their origin, but also on the starter culture used in the fermentative process.

A superior amount of nitrogen compounds leads to selecting these yeasts for industrial

purposes, but also to appreciating them from the point of view of yeast yealds obtained.

The study can offer alternatives when it comes to selecting these industrial yeasts, as the

purpose of their use matches the amount of nitrogenous substances determined.



130

Although the values determined in the present study appear to be similar, this is important at

the level of macro or microelements, as, qualitywise, it is a distinguishing factor between

yeasts.

Acknowledgement: This paper is supported by the Sectoral Operational Programme Human ResourcesDevelopment (SOP HRD), financed from the European Social Fund and by the Romanian Government under the contract number POSDRU/159/1.5/S/133675.

REFERENCES

1. Albers, E., C. Larsson, G. Lidén, C. Niklasson, and L. Gustafsson, (1996), Influence of the nitrogen

source on Saccharomyces cerevisiae anaerobic growth and product formation, Applied and Environmental Microbiology, vol. 62, no. 9, 3187–3195

2. Bell, S. J., Henscke, P. A., (2005) Implications of nitrogen nutrition for grapes, fermentation and wine, Australian Journal of Grape and Wine Research, Vol. 11, Issue 3, 242–295

3. Gardner, J. M., Poole, K.†, Jiranek,V., (2002), Practical significance of relative assimilable nitrogen requirements of yeast: a preliminary study of fermentation performance and liberation of H2S, Australian Journal of Grape and Wine Research, Vol. 8, Issue 3, 175–179

4. Mendez-Costabel, M.P. K.L. Wilkinson, S.E.P. Bastian, C. Jordans, M.McCarthy, C.M. Ford, N.K. Dokoozlian, (2014), Effect of increased irrigation and additional nitrogen fertilisation on the concentration of green aroma compounds in Vitis vinifera L. Merlot fruit and wine, Australian Journal of Grape and Wine Research, Vol. 20, Issue 1, 80–90

5. Oprean L., Dezsi C., Iancu R., Lengyel E.,(2012), Practical applications of yeast strans with superior biotechnological properties, Management of Sustainable Development, vol.IV,(1), 41-44

6. Rapp A and G. Versini, (1995), Influence of nitrogen compounds in grapes on aroma compounds of wines, Developments in Food Science, vol. 37, 1659–1694

7. Ribéreau-Gayon, P., D. Dubourdieu, B. Donèche, and A. Lonvaud, (2006), Handbook of Enology, John Wiley & Sons, 2nd edition

8. Tita, O., Oprean, L., Tita, M., Gaspar, E., Pacala, M., Lengyel, E., (2011), The influence of thiamine in the fermentation of the wine yeasts, Scientific Study and Research / Chemistry and chemical engineering, Biotechnology, Food Industry, 12(4), 381-386

9. Tiţa, O., Oprean, L., Tiţa, M-A., Păcală M., Lengyel, E., (2008), The influence of nitrogenous substances on the organoleptic characteristics of the wines during their evolution, Proceedings of the WAC 2008 International Conference, Beaune France, 255-257

10. Torrea, D., C. Varela, M. Ugliano, C. Ancin-Azpilicueta, I. Leigh Francis, and P. A. Henschke, (2011), Comparison of inorganic and organic nitrogen supplementation of grape juice—effect on volatile composition and aroma profile of a Chardonnay wine fermented with Saccharomyces cerevisiae yeast,” Food Chemistry, vol. 127, no. 3, 1072–1083

11. Varela, C., F. Pizarro, and E. Agosin, (2004), Biomass content governs fermentation rate in nitrogen-deficient wine musts, Applied and Environmental Microbiology, vol. 70, no. 6, 3392–3400



131

MOIST MATERIALS DRYING PROCESS ENERGY CONSUMPTION

CALCULATION Natalia Țislinscaia*, Mircea Bernic*, Andrei Lupaşco*, Alexei Buleandra**, Iana Ţislinscaia*,

Răducan Marcel*, Mihai Melenciuc*

*Technical Universiti of Moldova

**National University of Food Technologies, Kiev, Ukrain

Abstract: As the food moisture has different forms of communication, requiring large amounts of

energy for their destruction, we attempted to simulate the optimization of energy supply and to present it as a

function of the energy needed for evaporation depending on the energy needed to break ties between the

moisture and the product using internal heat source.

Key words: energy supply, drying process, optimization

Most foods contain a significant amount of water that enters the plant and animal

tissues and is a necessary part of them. However, an excess of water reduces the nutritional

value of food, increasing transportation costs and can cause damage to livelihoods as a result

of various microorganisms. In this regard, people must significantly draw attention to the

dried vegetables and fruits.

As the moisture in food has different forms of communication, requiring large amounts

of energy for their destruction, we attempted to simulate the optimization of energy supply

and to present it as a function depending on the energy needed for evaporation and the energy

needed to break ties between the moisture and the product i.e. internal heat source.

The basis of mathematical modeling method was based on Brandon method, which

allows us to obtain the nonlinear functional dependence of the output parameters and factors

of the process.

The work from a mole of water separation, for an isothermal reversible process is expressed

as:

lnln RT

Pp

RTAnas

p (1)

φ – (relative vapor pressure, is equal to the ratio of the vapor pressure of the material at this



132

moisture content U, to the vapor pressure of the water Pnas (the saturated vapor pressure at the

temperature T))

Differentiating (1) with respect to T, we are finding the total amount of heat,

indispensable for dehydration of the product.

01

2 )()lnln

( QuQTP

TP

RTQ nasp

(2)

Q1(u) - heat of evaporation of water from the material at this moisture content;

Q0 - evaporation heat of free water;

On the assumption of the equation (2)

TP

RTuQ par

ln)( 2

1 (3)

where R - gas constant of water vapor that is equal to 461.58 (J/kg K), T - temperature, K

Figure 1. The dependence of partial pressure on temperature for different humidities.

If substituting functional dependencies P =f(T) in the equation (3), we obtain the

calculated value Q1(u) - heat of water evaporation from the material at different moisture

content of the product:

TTRTQ

)16732551.55ln(%)5( 2

1

TTRTQ

)3306499.109ln(%)10( 2

1



133

TTRTQ

)4452488.148ln(%)20( 2

1

TTRTQ

)7650142.256ln(%)30( 2

1

TTRTQ

)9609286.322ln(%)50( 2

1

TTRTQ

)12019552.407ln(%)70( 2

1

Figure 2. The Heat of evaporation of water from the material.

The second constituent of the equation (3) represents the heat of evaporation of free water:

TPRTQ nas

ln2

0 (4 )

Herewith, this schedule of values of saturated vapor can be rather well approximated with the

formula: 1546.32 )273(1096.4 TPnas , Pа (5)

TTRTQ

))273(1096.4ln( 1546.32

20

(6)



134

Figure 3. The evaporation heat of free water.

Figure 4. Maximal balanced quantity of heat, necessary for evaporation of moisture from the material

and evaporation of free water.

Figure 4 shows the maximal balanced quantity of heat, necessary for evaporation of

moisture from the material and evaporation of free water. Binding energy decreases

dramatically with diminution of moisture content of the product. The difference of energy

consumption for the moisture of 5 and 70 percent, at 313 K consists 1372 J/kg, with

temperature increasing this difference considerably reduces and at 373K is approximately 70

J/kg.

Mathematical relation between Q and temperature for different humidities, was calculated

and presented by us as follows:



135

A received balanced functional relationship will form a base of the mathematical model

for optimization a power conduit with internal source of heat of tension of the

electromagnetic field, of the moisture, of electro physical operation factors - the dielectric

loss tangent and inductive capacity, and also the time of heating.

)()()()()()( 654321 fftgfTfWfEfQv (7)

Mathematical relationship (8) is based on non-linear forms of links of outside factors

xi, the solution of such models is possible using the method of Brandon [2]. For a more

accurate construction of functional dependence of the function fi(Xi) in expression (7),

should be placed in descending order the influence of Xi on , that is, to use ranking factors.

Derived functional dependencies allowed us to elaborate the model (7) - optimal

energy supply of the internal source of heat as a function of intensity of electromagnetic field,

of humidity, of electrical parameters - tangent of the angle of dielectric loss and permittivity,

as well as a heating time. This mate model takes into account a nonlinear functional

dependence of main ranged factors from the basic parameter.

(8)

The check on the adequacy of the mathematical model is studying using statistical

Fisher criterion [3].

Where,

Q - represents the energy required to carry out the process , KJ/k

W - humidity, %

Т - temperature, 0С



136

F - electromagnetic field frequency

- relative dielectric permittivity

tgδ - tangent of dielectric loss angle

Е - electromagnetic field strength, W/m

t - time ,s

RFERENCES

1. Лыков A.B. Теория сушки.-М.:Энергия,1968.-471с.

2. Бондарь Л.П. Математическое планирование в химической технологии. К.: В. школа, 1973 -

280с.

3. Грачев Ю.П. Математические методы планирования экспериментов М.: Пищевая

промышленность, 1979 - 200 с.



137

DRYING KINETICS OF ALMOND SEEDS USING PULSED UHF

ENERGY INTAKE

Mircea Bernic*, Natalia Ţislinscaia*, Lupaşco Andrei*, Vasile Cartofeanu*, Marcel Răducan*,

Leonid Zavialov**, Mihai Melenciuc M.*

*Technical Universiti of Moldova

**National University of Food Technologies, Kiev, Ukrain

Abstract: In this paper the drying kinetics of almond seeds using pulsed UHF energy intake are

given. It is shown and argued that the application of pulsed internal heat source made possible the

reduction of heat treatment period by 19,6%, and of specific energy consumption by 21 %.

Keywords: Drying, almond, UHF field, mass and heat transfer, pulse.

Introduction

As wrote in National Program for Nut Crops Developing in 2020 in Moldavia [1] are

planned to be planted 10,1 thousand hectares of almond orchards containing homologated

autochthones varieties, which will present a high adaptability level to our country south and

central regions and a harvest of 2,0 – 2,5 tones of dried fruits per hectare, within their full

fruiting period.

Of special importance, for our national economy, is the profound processing of that

production that allow us to transfer it from row material category to finished product one.

For storage terms augmentation, as well as for processing study, one of the key

operations is – drying, that usual is a long lasting process and demands high energy costs.

Drying intensification, which would also lead to energy costs diminution, is easy achieved by

augmenting drying agent temperature – using convection, or drying product temperature

while using UHF [2, 3]. Knowing that almond nuts are characterized by a high content of

fatty acids, possessing a superior nutritive value, which are extremely sensitive to high

temperatures, those kinds of drying augmentation aren’t the right one.

Mass and heat transfer phenomena research showed the possibility of reducing drying

time and energy costs using pulsed electromagnetic field. This, allows at product relative low



138

temperatures, a considerable augmentation, of one of the drying process transfer phenomenon

driving forces, namely – temperature gradient (T).


In the paper is presented a study of almond seeds drying kinetics, by applying internal

heat source – the ultra-high frequency electromagnetic field (UHF). Energy intake was

studied continuously and periodically – pulses containing of active heating period and passive

period of relaxation (lack of energy source).

For the study were used almond seeds of variety “Victoria”, with initial moisture of

26,89%. The seeds were calibrated to a further use of midsized ones (length 25±2 mm,

thickness – 8±1 mm).

Drying process was performed in a laboratory that allows the energy intake in various

ways: convection, UFH and combined (convection + UHF). Mass decrease was registered

online during drying process, using electronic scales SC-132 (cap. 300 g, div, 0,01g),

temperature of drying agent – with thermocouples, temperature at product’s center and

surface – with resistance thermometer NcW-1.617.

According to the obtained parameters there were obtained drying curves, drying speed

curves, average temperature dependence and variation of temperature gradient as a function

of time.


In order to study the effectiveness of the electromagnetic field applying during

almond seeds drying process, there was investigated drying kinetics with UHF energy intake

in both continuous and impulse regimes.

To ensure the pulsed regime there were determined formulas to calculate the duration

of active (energy intake) and passive (resting) periods of a pulse as a function of drying

parameters: field strength, temperature of product and its electro- and thermo-physical

properties [4].

MSVV

A TTQ

dcTQ

xc

2

, (1)

ppP a

dax

82

22

, (2)



139

where a – represents the active pulse’s period, s; – heat transfer coefficient, W/m2.K; –

thermal conductibility coefficient, W/m.K; d – characteristic product’s dimension, m; QV –

internal heat source’s strength, W/m3; TS – product’s surface temperature, K; TM –

environment temperature, K; p – passive pulse’s period, s; d – product layer’s thickness, m;

ap – molar diffusion coefficient, m2/s.

Pulsed UHF energy input with pulse’s parameters obtained by formulas (1) and (2)

serves to maintain the maximum values of the temperature gradient through the drying

period.

The drying kinetics was studied for the case of drying agent temperature of 20 °C, at

which in case of continuously UHF energy intake was found minimum energy consumption

compared to higher temperatures.

Field strength was calculated using formula (3) [5].

RRkf

TTE MC

110555.0 6. (3)

where TC – represents the temperature of the product’s center, °C; TM – environment’s

temperature, °C; R – product layer’s thickness, m.

The calculation of electric field intensity was performed in recitals as Tmax to be

obtained at the average temperature of 93 °C, temperature which is recommended by US

Department of Agriculture in collaboration with Almond Manufacturers Association of

California [5], and which assures the maximum extraction of essential oil without quality

thermal damage.

To maintain maximum values of temperature gradient in this regime, according to

formula (3) the electromagnetic field’s strength was accepted – 44,0 kV/m.

Drying curves W = f(τ) of almond seeds for the case of pulsed energy intake (curve 1)

and continuous energy intake (curve 2) are presented in fig. 1. Overall graphic character of

drying curves for pulsed energy intake completely corresponds to drying curves for

continuous one, except for a small deviation in time. This confirms once again the theory of

such scientists as A.V. Lycov, A.S. Ginsburg, V.V. Krasnicov, M.A. Grishin etc. [2, 3] that

the curves’ character largely depends on the physic-mechanical structure of the product and

on the related forms of moisture with the skeleton. However, on the drying curves are well



140

highlighted periods of UHF energy intake (slope with an advanced inclination angle) and

periods of relaxation (slope with a low inclination angle).

From graphs is observed that at pulsed energy intake compared to continuous energy

intake, drying period decreased by 19,6% being 118,3 min.

Fig. 1. Drying and temperature curves of almond seeds drying process with pulsed (curves 1

and 1`) and continuous (curves 2 and 2`) UHF energy intake

On the drying curves for pulsed UHF energy intake are well highlighted some waves

(curve enlarged in a circle) with the appearance of extremes. This indicates that when

disconnecting power supply, occurs a redistribution of mass and heat transfer resistance

forms, reducing them, which also favors the moisture transfer.

The same fig. 1 also shows the temperature variation character of almond seeds during

the drying process (curves 1' and 2'). As shown in the graphs for both cases of energy intake

(continuous and pulse), drying period increasing involves a temperature increase achieving

some maximum value.

The value of maximum obtained temperature is a function dependent on field

strength, electro- and thermo-physical properties of the product and energy application

method. For the case of continuous UHF energy input at field intensity of 44,0 kV/m was

obtained product’s temperature of 91,3 °C (curve 2'). For the case of pulsed UHF field intake

maximum values of the temperature was 85,2 °C (curve 1').



141

Duration of product temperature increase up to the maximum was 34 min. Further, for

both methods of energy input was maintained a relatively constant product temperature, all

the energy being consumed for the vaporization of free moisture and overcoming its moving

resistances from the center to the exterior layers. However, during continuous energy

application, it can be observed a temperature decrease of about 5% (curve 2`). Product’s

temperature decrease, at continuous heating, is caused by energy consumption increasing at

chemical and mechanical bonds breaking (between moisture and product’s skeleton) and

reducing the released in the product heat amount, as a consequence of polar water molecules

number reducing, which is mentioned in literature [6]. The phenomenon of product’s

temperature decrease in the second drying period practically disappears if using pulsed

heating. This is due to uniform moisture redistribution during relaxation.

One of the basic driving forces of the drying process is the temperature gradient. fig. 2

illustrates its variation during the drying process, using pulsed (curve 1) and continuous

(curve 2) energy intake.

Fig. 2. Temperature gradient variation during the drying process using pulsed (curve 1) and

continuous (curve 2) UHF energy intake

The graphs show that during continuous UHF energy intake, the temperature gradient

for E = 44,0 kV/m increases to 5,4 °C/m and then decreases to approx. 0,5 °C/m (curve 2).

Application of pulsed internal heat source from the moment the temperature gradient reaches

maximum values, allowed the maintenance of advanced gradient’s values throughout the

drying process (curve 1). Thus, if at continuous UHF energy intake, the maximum speed of

temperature gradient decrease was of 0,22 °C/(m.min), then at pulsed energy intake – only

0,074 °C/(m.min). So the temperature gradient, during drying process, was maintained about

2,9 times higher.

Fig. 3 presents the variation of active (energy intake) and passive (resting) period of

one pulse of UHF energy intake, depending on drying process duration. From the graphs is



142

observed that for the case of electromagnetic field’s intensity maintaining (44,0 kV/m), both

active period and the passive one are growing during drying. The duration of active period

grows faster than the duration of the passive one.

The increasing of the active (energy intake) period of the pulse during drying process is

a result of moisture content reducing, and therefore the amount of heat released in a unit of

time under the action of the electromagnetic field. Passive period (relaxation) increases due to

products thermal conductibility reduction at dehydration [7].

Fig. 3. Variation of UHF energy intake period (activ) and resting period (pasiv) of a pulse

during almond seeds drying using pulsed UHF energy supply

Application of pulsed electromagnetic energy fields have reduced the total duration of

heat treatment of almond seeds using UHF field of 0,73 times, or 61 min (brief duration of all

periods of rest during impulse drying).

Fig. 4 presents the curves of almond seeds drying speed using pulsed UHF energy

intake (curve 1) and continuous (curve 3). Curve 2 is the regression of curve 1.



143

Fig. 4. Drying rate curves of almond seeds: 1 – pulsed UHF energy intake, 2 – regression of

function 1, 3 – Continuous UHF energy intake

From the figure is observed that the drying rate curve for the pulsed UHF energy

intake (curve 1) draws the same shape as the drying rate curve for the continuous UHF field

intake, but has an oscillating character. Maximum extremes of the oscillations (pulse)

correspond to the period of active energy intake and minimum extremes - to the relaxation

period. Thus, the maximum drying rate for the active periods of energy intake for almond

seeds was 0,219 %/min and passive ones – 0,716 %/min. Compared to maximum drying

speed, at continuous energy intake was observed an increase of 24% in active periods and a

decrease of 25% during passive ones.

Comparative analysis of specific energy consumption for oil products drying using

pulsed UHF pulse energy intake comparative to continuous energy intake showed a decrease

from 0,76 kW/kg to 0,51 kW/kg, which consists 21,8%.

These energy consumption reductions are caused by drying time decreasing at the

same field strength, dehydration which takes place also in relaxation periods, by consuming

the cumulative energy and a better use of inertia effects, as well as because of the high values

of temperature gradient which reduce moisture transfer resistance through the capillaries.

However, energy consumption is reduced accordingly and the partial removal of moisture

from product’s surface, under the action of the air flow’s kinetic energy.

Conclusion

The study of almond drying kinetics has confirmed the application of pulsed internal

heat source. This made it possible to shorten the heat treatment product’s duration by 61 min

(by 19,6%) compared to continuous energy input. Reducing the duration of heat treatment

duration allowed a significant reduction of specific energy consumption by 21%.



144

Application of pulsed UHF energy showed a reduction of 5% of product’s average

temperature at the same field strength 44 kV/m.

Reduction of product temperature duration of heat treatment will influence the quality

indices of nutrients that are found in almond seeds, especially in fatty acids.

References 1. Programul Naţional Pentru Dezvoltarea Culturilor Nucifere Pină in anul 2020. Hotărâre de

Guvern nr. 8 din 03.01.06, Monitorul Oficial nr.005 din 13.01.06;

2. Лыков А.В. Теория сушки. – М.: Энергия, 1968. 470 с.

3. Малежик И.Ф., Тарлев В.П., Лупашко А.С. Конвективно- высокочастотная сушка

косточковых фруктов. Кишинев: UTM, 2005. - 460 c.;

4. Bernic M. Theoretical and experimental suppositions regarding the drying process of oily products

when using impulse heating sourse. / Buletinul Institutului politehnic din Iasi, Tomul LVI (LX)

Fasc. 4B, Ed: Politehnica, Iaşi 2010, p.341-346.

5. Hughes M.K., Yanamala S., San Francisco M., Loneragan G.H., Miller M.F., Brashears, M.M.

Reduction of Multidrug-Resistant and Drug-Susceptible Salmonella in Ground Beef and Freshly

Harvested Beef Briskets after Exposure to Commonly Used Industry Antimicrobial Interventions.

// Journal of Food Protection, Vol. 73, Num. 7, 2010 , pp. 1231-1237(7)

6. Hussain, A.; Li, Z.; Ramanah, D. R.; Niamnuy, C.; Raghavan, G. S. V. Microwave drying of

ginger by online aroma monitoring. / Drying technology, Vol. 28 Issue 1, 2010, p42-48,

7. Гинзбург А. С., Савина И. М. Массовлагообменные характеристики пищевых продуктов.

Справочник. - М.: Легкая и пищевая промышленность, 1982. – 280с.



145

DETERMINATION OF RELATIVE SPEED OF PARTICLES IN

LIQUID FLOWING IN TUBES AT HIGH TEMPERATURES

C. Căpăţână *, M. Văduva *

* "Lucian Blaga" University Sibiu, Faculty de A.S.F.I.E.P.

A laboratory scale apparatus was designed to carry out studies on the flow behavior of particles suspended in

carrier fluids at elevated temperatures. Single particles were introduced into a glass tube to simulate

conditions existing in holding tubes of continuous flow systems. Tests were conducted with water and 3%

starch solutions at temperatures of 100oC, over a range of fluid flow rates. Food particles (apple and pear

cubes) as well as polypropylene spheres were used to test the suitability of the apparatus.

Keywords: relative speed particles/fluid; particle speed, flowing.

Introduction

Particle behavior in tubes determine the design and development of the equipment for

aseptic processing in order to ensure the quality and safety of food products such as mixes of

particles and liquids.

Models shows that the size of particles, the resting time and the heat transfer

coefficient are important in quality evaluation and in sterilization process. The particle speed

depends on size, fluid viscosity and tube geometry.

Materials and methodes

Experimental instalation consist of:

- a jacketed tank;

- a pump for warm liquids;

- a flood gate for solid particles intake;

- a glass tube for visualization.



146

Jacketed tankHeated liquid

Pump

Flood gate for solidparticles intake

Rubber sleeve Transparent glass tube

Drain connection

Figure 1. Experimental instalation for studying particles speed.

Fluid temperature is measured with a thermometer placed in liquid. Liquid pump can offer

various flow rates. The solid particles are introduced one by one in tube in order to observ

their behavior. The glass tube is mounted with two rubber sleeves at the ends.

Experimental

Experiments are made in water and starch solution both at 100oC. Polypropylene

spheres have 0.635, 0.953, 1.27 and 1.9 cm diameter and apples and pears diameters are 0.75

and 1.27 cm.

Fluid flow rate was determined by volume measurements in fixed periods of time.

Results and discution

The temperature of 100oC is important to study particles behavior in such conditions

near industrial processing. Raw data is presented in Table1. Table 1

Transp.

fluid

Particles size (cm)

Polypropylene Apple cube Pear cube

0,64 0,95 1,27 1,91 0,76 1,27 0,76 1,27

Starch solution 3% at flow rate 13 l/min and average speed of 0,161 m/s

VMP* 0,074 0,103 0,125 0,155 0,156 0,194 0,140 0,180

VR* 0,46 0,64 0,77 0,96 0,97 1,21 0,89 1,12

Water, flow rate at 13 l/min, average speed of 0,158 m/s

VMP* 0,121 0,133 0,136 0,148 0,144 0,149 0,128 0,121

VR* 0,77 0,84 0,86 0,94 0,91 0,95 0,81 0,77

*VMP – average speed of particles (m/s);

VR – relative speed (particles speed/fluid speed)



147

The results indicate that at a 13 l/min flow rate bigger size particles crosses the tube

faster than smaller particles. We observed that particles can achieve faster speed than the

transporting fluid. In water speed profile is the same but in starch solution the relative speed

of food products is subunitary. This behavior is due to viscosity and density differences,

which induce a lifting effect to particles. Another fact that affect the speed of polypropylene

particles is the friction between particles and tube inner walls.

Generaly the relative speed rise with particles size. In starch solution food particles

speed is faster than transporting fluid relative speed at higher diameters.

In figure 2 is presented variation of particles speed for various sizes at 13 l/min flow

rate and 100oC temperature.

Particles speed in water

0

0,05

0,1

0,15

0,2

0,635 0,953 1,27 1,9Particles diameter(cm)

Parti

cles

spe

ed(m

/s)

polypropileneapplepear

Particles speed in starch solution 3%

0

0,05

0,1

0,15

0,2

0,635 0,953 1,27 1,9

Particles diameter(cm)

Parti

cles

spe

ed(m

/s)

polypropyleneapplepear

Figure 2. Particles speed variation for different sizes.

References

Căpățână C. și Văduva M.: „Operații unitare în ingineria alimentară”, Ed. Universității „Lucian Blaga” Sibiu,

Sibiu, 2010.

Răşenescu Ion : “Operaţii şi utilaje în industria alimentară”, Vol.1, Ed. Tehnică, Bucureşti, 1971.

Bratu E.A. : "Operaţii unitare în ingineria chimică", Vol.1, Ed. Tehnică, Bucureşti, 1984.



148

REDUCEING HUMIDITY LOSSES OF FRUIT PIECES BY COATING

WITH BEESWAX

C. Căpăţână *

* "Lucian Blaga" University Sibiu, Faculty of A.S.F.I.E.P.

Fresh whole peeled fruits or segments are considered minimally procesed products. When dry

packed, segments from mature fruits had fluid losses 15% during 4 wk storage. Edible wax

microemulsion coatings reduce leakage of dry-packed segments by 80% after 2 wk, and 64% after 4 wk

storage, and were optimally effective in reducing leakage when diluted to 12% solids. Coatings could be

made with polyethilene, candelilla wax, with lauric, stearic, palmitic, oleic or myristic acids. Coatings

were not detected by informal taste panels before or after storage.

Keywords: fruit coatings, beeswax solution, humidity loss.

Introduction

Fresh whole or pieces of fruits are sold in their natural state, unpreserved. Lifetime of

such product is usually between 17 and 21 days, depending on storage condition.

Along flavor loss and microbial spoilage another key factor limiting lifetime is water

loss. When semnificative water loss fruits become unsoldable. In order to preserve water

inside fruits there were attempts to cover their surface with an edible coating made of calcium

aliginat.

This type of coating extends resistance of fruit surface but the effects on water loss

was neglectable. An increase about 89% in surface resistance lead to only 16% reduceing of

water loss, due to hydrophilic properties of calcium aliginat membrane which cannot prevent

the migration of water from inside to outside of fruits.

The humidity variation during marketing seasons is shown in figure 1.



149

Humidity loss

0

510

15

20

NOV DEC JAN FEB MAR APR MAY

MonthLi

quid

loss

(%)

Figure 1. Seasonal humidity loss of fruits at 2oC

Experimental

Aplying of beeswax was made with a brush or by dipping fruit pieces into wax

solution.

Beeswax solutions were made at the right solid concentration. Fruit pieces were

dipped for 30 seconds, then the excess was removed by gravitational drain over a plastic grid.

There were only a few points of contact between fruit and the grid. Later there was a slow

ventilation applyed to fruit pieces in order to dry the wax. For dipped pieces drying take 150

minutes and for brushed pieces 60 min.

Each lot was placed into polyethylene bags and stored at 2oC. Humidity losses was

determined for 4 weeks by measuring weight.

If the wax is well spred there is no affected taste along with the fluid loss reduction.

Senzorial analyze shows that nor after one day neither 3 weeks the wax coating didn’t affect

fruit flavor or taste, comparing with fresh samples.

The different type of coating is shown in figure 2, where we can observe that dipping

method is the best.



150

Coating Effect

0

10

20

30

40

0 1 2 3 4 5

Weeks

Hum

idity

loss

(%)

unprotectedbrushingdipping

Figure 2. Coating effect upon humidity losses during storage of fruit pieces

In order to determine the best solid concetration of beeswax solution there were several

values tried such as: 18, 15, 12, 9%. Humidity losses are strongly reduced at 12% beeswax

solution (figure 3).

Influence of beeswax concentration

05

1015202530

0 1 2 3 4 5Weeks

Hum

idity

loss

(%)

other9% s.u.12% s.u.

Figure 3. Humidity losses depending on beeswax solution concentration

Fatty acids from beeswax helps retaining water. Microemulsions of beeswax consist of wax,

a fatty acid and a base. As base was used ammonia, as fatty acids oleic, stearic and palmitic

acid.

To sumarize, beeswax solutions mixed with fatty acids (C12 to C18) can provide protection

against humidity losses, prolonged self life of fruits with almost 4 weeks. Optimal storage

condition is obtained useing 12% solid compound concentration. There are no modifications

regarding flavor or taste from the first to 21-th day of storage.



151

References 1. Baldwin E.A. şi Baker R.A. : "Use of edible coatings to preserve quality of lightly processed products", Food

Science and Nutrition 35(6) 1995.

2. Baldwin E.A. şi Nisperos-Carriedo M.O. : "Use of edible coatings for lightly processed fruits and

vegetables", HortScience, 30(1) 1995.

3. Ciprian Căpățână: Teză de doctorat „Studii și cercetari în domeniul ambalajelor comestibile folosite în

industria alimentară”, Sibiu, 2003;

4. Ciprian Căpățână: „Ambalarea produselor alimentare”, Ed. Universității „Lucian Blaga” din Sibiu, Sibiu,

2000.



152

EXTRACTION OF ANTHOCYANINS FROM BY-PRODUCTS

FROM THE WINEMAKING PROCESS

Adina FRUM*

*PhD Fellow, SOP HRD/159/1.5/S/133675 Project, Lucian Blaga University from Sibiu, Partener, e-mail:[email protected]

Abstract

Anthocyanins are polyphenolic pigments that can be found in berry fruits, cereals and vegetables.

They are responsible for colors like red, blue or purple found in nature and can be used as natural coloring

agents in pharmaceuticaland food industries. The samples used for anthocyanins extraction were pomaces

from Cabernet Sauvignon and FeteascaNeagra cultivars from Romanian wineries. For the identification of

the compounds the absorption spectrum was used and it had an absorption maximum at 535nm, consisting

with the presence of anthocyanins. The methods used for extracting anthocyanins from grape pomace were

using an ethanol and hydrochloric acid solvent, extracted for 36 hours at 4ºC, room temperature and after 30

minutes in the ultrasound bath at room temperature. The largest amount of anthocyanis was found in the

Cabernet Sauvignon cultivar pomace that was extracted at room temperature after ultrasound treatment. The

amount of anthocyanins extracted from the analyzed pomaces is adequate for usage in several industries like

food industry or pharmaceutical industry. Keywords:anthocyanins, natural pigments, extraction, pomace, ultrasounds, temperature

Introduction

In the winemaking process, besides the main product, there can be found some other

compounds that have proven to be useful, called by-products. By using these by-products the

pollution of the environment can be decreased and several compounds such as polyphenols,

enzymes and vitamins can be recovered [1].

Anthocyanins are polyphenolic pigments that can be found in berry fruits, cereals and

vegetables. They can provide colors like red, blue and purple found in nature [2]. They are

responsible for the red color of grapes and wine. This characteristic is determined by their

degree of hydroxylation, methylation or glucosidation [3].

The quantity of anthocyanidins in grapes is different due to the grape cultivar, the

degree of maturity at the harvesting moment and the weather conditions, especially the

temperature and the light conditions [3].



153

Most of the anthocyanins found in grapes are located inside the grape skins and only

a part of them are extracted into the wine due to the capacity of the cellular wall to degrade

using pectolytic enzymes, thus making cell wall pectins permeable and susceptible to the

changes that happen in the vinification process [4].

Anthocyanins are used in the food industry as a natural coloring agent for the acid

foods, in the pharmaceutical and cosmetical industry replacing the synthesis coloring agents

[5].

Over the years, several health benefits have been found that are possessed by these

compounds. They have antioxidant, anti-inflammatory, anti-ageing activities and they can

prevent cardiovascular diseases[2,6], diabetes[7], obesity[8] and they can be used as topical

agents for skin protection against UV radiation[9].


The samples used for this experiment were by-products of the winemaking process.

They consisted of red grape pomace: seeds, stems and skins from Cabernet Sauvignon and

FeteascaNeagra cultivars from Romanian wineries. The grape pomace was dried at a

temperature below 40ºC, ground on a domestic mill and stored at -20ºC until use.

1 g of red grape pomace was extracted with 100 mL of ethanol and hydrochloric acid

(85:15) solvent for 36 hours, at 4ºC, room temperature and after 30 minutes in the ultrasound

bath at room temperature. The mixture was filtered, brought to 100 mL with ethanol and

hydrochloric acid (85:15) solvent and analyzed spectrophotometrically after a 1:2 dilution

with the same solvent.The absorption spectrumwas recorded in UV-VIS range and the

absorption was recorded at a wavelength of 535nm.

The specific absorption range of anthocyanins is 500-535 nm, meaning that the

substances that have an absorption maximum within that range are identified as anthocyanins.

The amount ofanthocyanins can be determined by measuring the absorption at a

wavelength from their specific absorption range.

The total amount of anthocyanins was determined using the following formula:

(1)

Where:

E= absorption of the solution at 535 nm

d=dilution factor (in mL)



154

98,2= absorption value of the solvent ethanol and hydrochloric acid (85:15), at 535

nm


Using the absorption spectrum (Fig. 1,2), from the spectrum curve, there can be seen

that the absorption maximum is at 535 nm, characteristic to anthocyanins. All the samples

had the same absorption maximum that was consisting with anthocyanins, so all the samples

contained anthocyanins.

Fig.1 Cabernet Sauvignon samples absorption spectrum

Fig.2 FeteascaNeagra samples absorption spectrum

Using formula (1), the amounts of anthocyanins were calculated and then represented

in a chart (Fig. 3).



155

Results show that the pomace from the Cabernet Sauvignon cultivar had a greater

amount of anthocyanins than the pomace from theFeteascaNeagra cultivar.

The quantity of anthocyanins differs not only because of the difference in cultivars,

but also because of other factors such as geographic region, agronomic factors and climatic

factors.

The extraction method has a great influence upon the amount of extracted

anthocyanins as well (Fig. 3 ).

Fig. 3 Extraction of the anthocyanins from the FeteascaNeagra and Cabernet Sauvignon pomace

The most effective extraction method was the extraction using the ultrasound bath. In

both cultivars the samples that were treated with ultrasounds showed a greater amount of

anthocyanins than the other samples.

The samples that were kept at 4ºC, showed more anthocyanins than the samples that

were kept at room temperature. That means that anthocyanins can be extracted better by

using a lower temperature.

Conclusions

Based on the absorbtion spectrum, the samples of pomace from the Cabernet

Sauvignon cultivar and FeteascaNeagra cultivar contained anthocyanins because of the

absorbtionmaximum that has been measured at 535nm, wavelength specific to anthocyanins

that is between 530 and 535 nm.



156

The amount of anthocyanins found in pomace from the Cabernet Sauvignon cultivar

was greater than the pomace from the FeteascaNeagra cultivar not only because of the

difference in cultivars, but also because of other factors such as geographic region,

agronomic factors and climatic factors.

The largest amount of anthocyanis was extracted after exposure to ultrasounds for 30

minutes and then extracted at room temperature even though the amount of anthocyanins

extracted at 4ºC was greater than the one extracted at room temperature. Ultrasounds allowed

the samples to have a more intimate contact with the solvents, thus the amount of

anthocyanins extracted was grater.

The amount of anthocyanins extracted from the analyzed pomaces is adequate for

usage in several industries like food industry or pharmaceutical industry as natural coloring

agents that replace the synthesis ones or because of their benefits in healthcare.

Acknowledgements:This paper is supported by the Sectoral Operational Programme Human

Resources Development (SOP HRD), financed from the European Social Fund and by the Romanian


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